The Qatar International Conference on Stem Cell Science and Policy

Breast cancer is a major disease leading in both incidence and mortality in women. Although the mortality has been reduced by various therapy approaches, recurrence still occurs in many patients. Evidence indicates that tumor-initiating (cancer stem) cells may contribute to disease relapse, suggesting the importance of effective targeting of this cell population. Recently, it has been shown that the components of the tumor microenvironment, including tumor vascular endothelium, interact with cancer stem cells through intracellular signaling pathways. However, the exact mechanism underlying this interaction is not known. In this study, we intend to investigate how the Akt-activated vascular endothelial cells (E4ORF1) communicate with defined populations of breast tumor. Moreover, we are interested to find out if notch pathway is regulating this crosstalk.

The cancer stem cells were enriched under serum-free conditions in 3D media to obtain mammospheres. To investigate how E4ORF1 maintains breast tumor cells and cancer stem cell in the absence of serum and cytokines, breast cancer MDA-MB231 and MCF-7 cells were co-cultivated with E4ORF1, its conditioned media, or in 3D media. In addition, the expression of pluripotency markers was assessed by real-time PCR in cancer cells after contact with E4ORF1 cells. Besides, the involvement of notch pathway in E4ORF1 interaction with mammospheres was studied by using RNAi against Jagged-1.

Our preliminary results demonstrated that E4ORF1 cells were able to maintain cancer cells under serum-free conditions and in a contact-dependent manner. Also,they were able to enrich mammospheres in contact or after temporary exposure. Moreover, the expression of some pluripotency markers displayed a 1.5-fold increase in cancer cells co-cultured with E4ORF1 cells. Besides, Notch pathway activation seemed to be necessary for E4ORF1 to support mammosphere growth since there was a 3-fold decrease in sphere numbers in Jag-1-/MDA-MB231 cells.

These results suggest that E4ORF1 cells are capable of maintaining cancer stem cell population. Since this interaction is contact dependent, it might indicate the involvement of some intracellular signaling pathways. Our results show that notch might be involved in this crosstalk, which makes it a target for therapy in order to inhibit the cancer stem cell population to prevent breast cancer reappearance.

In vitro neural differentiation of human embryonic stem cells (hESC), or induced pluripotent stem cells (iPSCs) is a powerful system to study early neural development, instrumental for cell replacement therapy and neuropharmacological studies. Here we report the derivation of tyrosine hydroxylase (TH) positive cells from human iPSCs, indicating the differentiation into dopaminergic neurons.

Our research focuses on understanding how pluripotent mammalian stem cells maintain their undifferentiated state and undergo differentiation in culture – this reflecting my enduring interest in the emergence of diversity during mouse gastrulation.

In recent studies we have examined the role of transforming growth factor family members in both pluripotency and differentiation. This involved analysis of the signalling cascade induced by treating hESCs with Activin or Nodal, determining how their response to these growth factors maintains hESC pluripotency. We started by developing chemically defined culture conditions in which the activities of specific growth factors could be identified and studied in a controlled manner. We then carried out a detailed analysis of the roles of Smad proteins (Smad2 and Smad3) as direct regulators of Nanog, which in turn blocks an hESC default differentiation into neuroectoderm. These studies led to our discovery of a novel type of pluripotent epiblast stem cell (EpiSC) from the late epiblast layer of mouse and rat embryos. EpiSCs share many features with hESCs, and subsequent work supports our hypothesis that hESCs are the human counterparts of EpiSCs, with similar responses to growth factors and mechanisms of pluripotency and differentiation.

In our most recent work we have focused on the role of bone morphogenetic protein (BMP)-4 in the cell fate decision between endoderm and mesoderm, demonstrating the similarity of BMP-induced hESC and EpiSC differentiation to mesoderm induction during mouse gastrulation. This work reveals the importance of BRACHYURY and CDX2 genes as key mediators of embryonic and extraembryonic lineage differentiation in hESCs and EpiSCs. Our focus on mesoderm leads us on to molecular pathways for early human cardiomyocyte differentiation, with a goal of understanding the transcriptional networks responsible for cardiomyocyte identity and using this to generate more homogeneous cardiomyocyte populations for therapeutic applications and drug discovery. Taken together, these studies should significantly accelerate the progression from basic stem cell research to clinical applications.

We have previously shown that Endothelial Progenitor Cells (EPCs) bind and inhibit platelet function and impair thrombus formation, but the surrounding mechanisms that regulate this process have not been fully described. Herein, we addressed the mechanistic action of EPCs on platelet-thrombus formation and we highlighted the role of platelet P-selectin in this process.

EPCs were generated from human peripheral blood mononuclear cells after 10 days of culture on fibronectin in conditioned media. The impact of EPCs on platelet aggregation and thrombus formation was investigated in P-selectin deficient (P-sel-/-) and their wild-type counterpart (WT) mice. EPCs significantly impaired, in a concentration dependent-manner, collagen-induced whole blood platelet aggregation in WT mice; whereas in P-sel-/- mice, EPCs had no significant effect. Moreover, in ferric chloride-injured arterial thrombosis model, infusion of EPCs significantly reduced thrombus formation in WT, but not in P-sel-/- mice. Furthermore, the number of EPCs recruited within the thrombi and along the arterial wall was reduced in P-sel-/- mice as compared to WT mice, and the relative mass of thrombi generated in EPC-treated P-sel-/- mice was significantly larger than that in EPC-treated WT mice.

EPCs impair platelet aggregation and reduce thrombus formation by a cellular mechanism implying binding to platelet P-selectin. These findings add new insights into the role of EPCs in the regulation of platelet function and thrombotic reaction during vascular repair.

Heart disease is a leading cause of death in adults and children. We, and others, have described complex signaling, transcriptional and translational networks that guide early differentiation of cardiac progenitors and later morphogenetic events during cardiogenesis. We found that networks of transcription factors and miRNAs function through positive and negative feedback loops to reinforce differentiation and proliferation decisions. Many of the same cues have been leveraged to control differentiation of pluripotent stem cells into cardiac, endothelial and smooth muscle cells that may be useful for regenerative purposes. We have used similar approaches to direct differentiation of disease-specific human induced pluripotent stem cells in order to model human heart disease.

Recently, we utilized a combination of major cardiac regulatory factors, Gata4/Mef2c/Tbx5, to induce direct reprogramming of cardiac fibroblasts into cardiomyocyte-like cells with global gene expression and electrical activity similar to cardiomyocytes. The reprogramming was a stable event at the epigenetic level. We have now used genetic lineage-tracing techniques to demonstrate that resident cardiac fibroblasts can be reprogrammed into induced cardiomyocytes in vivo using a gene therapy approach. The in vivo efficiency of reprogramming into cells that are more fully reprogrammed into beating cardiomyocytes was greater than in vitro and resulted in improved cardiac function after injury. Knowledge regarding the early steps of cardiac differentiation in vivo has led to effective strategies to generate necessary cardiac cell types for disease-modeling and regenerative approaches, and promise to lead to new strategies for human heart disease.

Since the first report on the generation of pluripotent embryonic stem cells (ESCs) from preimplantation embryos, there has been tremendous interest in developing culture systems to produce ESCs from human and other species efficiently to use them in biomedical applications. These cells have provided fascinating possibilities and tools to study human development, genetic diseases, and development of toxicological and pharmaceutical applications as well as in therapeutic transplantation. Developing an efficient method to derive ESCs from various genetic backgrounds should be valuable for establishment of ESCs in various mammalian species, including human. However, it is still encumbered by various technical and ethical objections. This challenge is circumvented by using small molecules that suppress differentiation pathways. Here, we will present our recent studies on efficient and reproducible establishment of ESCs from blastocysts and single blastomere to avoid destruction of the embryo. Additionally, we present our recent researches on testis-derived pluripotent stem cells that share characteristics of ESCs. The application of these small molecules will not only solve the problems of derivation and expansion of pluripotent stem cell in defined conditions, but also clarify new signaling pathways in them.

Human stem cells, particularly embryonic stem cells, have unlimited capacity to develop into different types of cells such as muscle cells, nerve cells, heart cells, blood cells and others. Therefore, human stem cells hold the promise of a renewable source of replacement cells and tissues to treat an ever growing list of human diseases and conditions, including Parkinson's disease, amyotrophic lateral sclerosis, spinal cord injury, burns, heart disease, diabetes and arthritis.

On the other hand, stem cell research has led to great controversy. The controversy centers on the ethics of conducting research involving the generation, usage and destruction of human embryonic stem cells. But it should be noted that new approaches and innovative strategies have yielded human stem cells without destruction of embryos via generating pluripotent stem cells from adult cells.

The Supreme Council of Health (SCH) has developed a policy for the ethical use of human stem cells in research. The policy provides guidance and procedures under which proposed research in this area, conducted in Qatar publicly or privately funded, is ethically responsible, scientifically worthy, and conducted in accordance with national and international laws.

The SCH presentation will introduce the research community to what is allowed and not allowed in research involving human embryonic stem cells in Qatar.

The low incidence of CFU-F significantly complicates the isolation of homogeneous populations of MSC from mouse BM (mBM), a common problem being contamination with hematopoietic cells. Taking advantage of burgeoning evidence demonstrating the perivascular location of MSC, we hypothesized that a potential reason for the low yield of mMSC from mBM is the flushing of the marrow used to remove single cells suspensions and the consequent destruction of the marrow vasculature, which may adversely affect recovery of MSC physically associated with the abluminal surface of blood vessels.

Herein, we describe a simple methodology based on preparation and enzymatic disaggregation of intact marrow plugs that yields distinct populations of both stromal and endothelial cells. The recovery of CFU-F obtained by pooling the product of each digestion (1917.95+199) reproducibly exceeds that obtained using the standard BM flushing technique (14.32+1.9) by at least two orders of magnitude (P<0.001; N = 8) with an accompanying 196-fold enrichment of CFU-F frequency. Purified BM stromal cell populations devoid of hematopoietic contamination are readily obtained by FACS at P0 and from freshly prepared single cells suspensions. Furthermore, this population demonstrates robust multilineage differentiation using standard in vivo and in vitro bioassays.

Since the first CB transplant (CBT) was performed by Gluckman et al in 1988, >16,000 patients have received this procedure for a variety of malignant and non-malignant diseases. Despite the lower frequencies of graft versus host disease (GvHD) and event-free survival rates comparable to those achieved after unrelated allogeneic marrow transplantation, a major disadvantage of CBT is the low cell dose, which results in slower engraftment and an elevated risk of engraftment failure by comparison with marrow or PBPC transplantation, even following transplantation with two CB units. A significant shortcoming of the cytokine-driven suspension culture systems initially envisioned for ex vivo expansion of CB cells is their failure to incorporate cues provided by the stromal cell-mediated microenvironment of the bone marrow. Seeking to enhance the efficiency of these culture systems, we have therefore incorporated mesenchymal precursor cells (MPC), a key cellular constituent of the hematopoietic stem cell niche. Optimisation of the expansion culture system and validation of the expanded product in immunodeficient NOD/SCID/IL-2RγNull (NSG) mice led to a clinical trial using haploidentical MSC (from a family member) and more recently using immunoselected allogeneic MPC (Mesoblast Ltd). More than 30 patients with a range of hematological malignancies have now been transplanted with a combination of two CB units, one unmanipulated, the second expanded on MPC. In fulfillment of our hypothesis, a significant enhancement in the rapidity of neutrophil and platelet reconstitution was observed, derived from the expanded unit that was superseded by long-term engraftment from the unmanipulated unit.

One of the fields of medicine that has raised the most expectations in Iran is cell therapy, using somatic or stem cells. The word “cell therapy” refers a series of procedures, which are used to separate and transplant cells in target tissues. However, in the absence of a current consensus on a approved protocol to isolation and identify suitable cells, the most methods in this field are under research yet. Royan Institute, as a leading center in stem cell research, in Iran, simultaneous with global research and increasing the knowledge about stem cells, the first Royan clinical trial in cell therapy field was done in 2004, which was designed and implemented for application of stem cells in patients with recent heart attack. During implementation of this project, at least three other projects were added to the list of Royan institute clinical trials. Two years later, Royan institute started to established GMP facilities for cell culture. By the end of year 2008, increasing number of clinical trials from 4 to 10 studies and termination of investigating stages of using Melanocytes for treatment of vitilligo disease and Limbal stem cells culture for using in patients with Limbal disease, led to establishment of a center for providing medical services to the patients. Now Royan institute in Regenerative department are doing more than 20 NIH registered clinical trials and there are some result in heart, bone, cartilage, limbal, liver and skin field, which will be presented in this lecture.

Haematopoietic stem cell transplantation (HSCT) is the treatment of choice for patients with refractory leukemia. However, HSCT is associated with lethal GVHD and/or disease relapse. The addition of alternative remedies such as adoptive immunotherapy may lead to eradication of minimal residual leukemia and prevention of relapse. Antigen-pulsed ex-vivo dendritic cells (DCs) have been utilized for the initiation of such immunotherapeutic modalities. DCs can successfully be generated from CD34+ haematopoietic stem cells (HPSCs) and DCs generated from patients with chronic myeloid leukemia (CML), displayed clonal heterogeneity with respect to the expression of the bcr/abl fusion gene. One reason could be that, DCs mixtures were originated from diverse sources of normal and leukemia either CD34+ or CD34- HPSC. CD34-HPSCs were reported to possess hematopoisis reconstitution capabilities. So far, the differentiation of DCs from CD34- HPSCs and their expression of bcr/abl have not been explored in CML patients. Consequently, we hypothesized that CD34-HPSCs could be a valuable source for the generation of an effective vaccine which could be stored and used latter to target residual leukemia if the patient encountered any relapses.

Our study showed that DCs generated from CD34-HPSCs were potent and efficient antigen presenting cells and comparable to their counterparts DCs from CD34+ HPSCs and hold promises in immunotherapy modality.

1A. Gaafar, 2H. M. Al-Omar, 1Z. Al-Mokhlafi, 1Manogaran PS, 1A. qniebi1, 1A Al-Mazrou, 2F.Al Mohareb, 1C. Adra, 1K. Al-hussein

1Stem Cells Therapy Program, 2 King Faisal Cancer Center, King Faisal Specialist Hospital & Research Centre.11211Riyadh, Saudi Arabia

Transplantation of allogeneic bone marrow has been successfully used in the treatment of high risk or recurrent hematologic malignancies, bone marrow failure syndromes, selected hereditary immunodeficiency states and metabolic disorders. Early in the history of bone marrow transplantation, it was clear that access to a suitable donor was a major obstacle severely limiting the use of this potentially curative treatment modality. Although using an HLA sibling donor is the best choice for transplantation, not all patients could have access to such a donor, therefore transplant centers explored the possibility of using volunteer adult unrelated donors as an alternative to HLA–matched siblings. In addition, there was another alternative treatment strategy as a source of hematopoietic stem cell namely Umbilical Cord Blood. In Saudi Arabia, 60 percent of our patients who need a transplant will find an HLA-matched sibling donor, leaving 40 percent of the patients in need of alternative sources. The concept of establishing a Cord Blood Bank in Saudi Arabia under the umbrella of King Faisal Specialist Hospital & Research Centre was raised after the increase in the rate of using cord blood for transplantation due to the inability of finding fully or closely HLA-matched related donors. This Cord Blood Bank is a non-profit public cord blood bank dedicated to making high quality cord blood units available to all patients in need of related and/or unrelated transplantation in the Kingdom of Saudi Arabia and in the neighboring countries through the development and maintenance of a center of excellence for the collection, storage, search and distribution of ethnically and racially diverse cord blood units. Additionally, the mission of the Cord Blood Bank is to educate both the medical community and the public to the value of cord blood donation collection and cryopreservation, and increase the awareness of the importance of cord blood banking. To date, our inventory consists of 3,725 units of high quality cord blood with a total of 70 cord blood transplants carried out from our own inventory.

This presentation will provide an additional perspective to that of the scientific and ethical issues that surround biosamples, by considering the appropriate role of regulation in this area. A regulatory regime can be used to ensure that the individuals, organisations and activities are governed in such a way as to ensure the highest ethical and scientific standards are maintained. Right touch regulation should promote innovation and should not be overly burdensome, but it is required to ensure that, in the highly sensitive area of biosampling, there is an appropriate framework in which high calibre operators can function and unsafe or unethical practise is prevented. As stem cell and biosample banking expands into new scientific and geographic areas regulation has to be sufficiently flexible to adapt to the unknown. It is also required if the bio banking communities are going to operate internationally.The presentation will consider what the international community might expect to see in regulatory terms as the Middle East expands into these opportunities. Regulation should be understood as key to ensuring confidence in the processes involved in biosampling. The presentation will look at some of the regulatory issues and challenges encountered in Europe and beyond and discuss the perspective and lessons to be learned. Central to the discussion will be consent issues and how consent may be embedded in regulation to underpin approaches to the storage, import and export and research with human tissue and cells.

Human stem cell research, because of its potential for both transformative sources of knowledge and for translation into widespread practical applications, has been a subject of ethical, religious, and policy debates since the first cells were characterized. The technology touches on some of humanities most urgent concerns and some of our most fundamental narratives. Stem cell research is characterized by a swiftly moving knowledge base, with first human embryonic (hES), then induced pluripotent cells (iPS) being seen as the best new hope in the search for cures to diseases. Each new advance in science brings new ethical and moral quandaries to the debate. This presentation will outline the history of ethical questions surrounding stem cell research, describe the variety of responses to such concerns, and define some of the international policy that has emerged to create ethical frameworks for ongoing research on stem cells. We will consider what questions, if any , can be understood as settled within a general consensus, and what questions may be raised by the future directions of stem cell research.

The use of donated eggs is an important component of assisted reproduction and plays a prominent role in certain lines of scientific inquiry, particularly in the area of stem cell research. Yet despite these benefits, egg donation remains extremely controversial. One result of this ethical controversy has been the development of a heterogeneous policy environment where the rules for egg donation for both reproduction and research vary substantially from one country to the next or even within countries. In this presentation, I review the major policy options for egg donation that have been adopted around the world and discuss the justifications for these policies. Then, drawing on international data on assisted reproduction and published stem cell research using donated eggs, I offer a preliminary assessment of how these policy choices affect the availability of donated eggs for both reproduction and research purposes.

There has been significant interest in the therapeutic and scientific potential of stem cells since reconstitution of the haematopoietic system was first realized by bone marrow transplantation in the 1960s. The isolation of tissue-specific, multipotent stem cells from adult organs and the derivation of pluripotent human embryonic stem cells offer the potential for regeneration of a number of different tissues and organs susceptible to age-related degenerative conditions and traumatic injury. In the not-too-distant future, it will be possible to repair heart tissue damaged by myocardial infarction, to replace neuronal cells lost in Parkinson’s and Alzheimer’s diseases, to transplant new insulin-producing cells for diabetics and myelinating cells for individuals afflicted with multiple sclerosis, and to replace bone and cartilage lost through aging and inflammatory disease. In addition, the generation of specific populations of defined subtypes of human cells has tremendous potential to revolutionize the fields of drug discovery and investigation into the cellular bases of human disease. The newly emerging field of regenerative medicine will fundamentally alter clinical medicine and significantly influence our perceptions of aging, health and disease, with a myriad of consequences for society at large.

Objective: To study efficacy and safety of autologous bone marrow stem cell therapy for tissue and neoangiogenesis in ischemic parts of heart in patients with coronary heart disease (CHD) and post-infarction cardiosclerosis after macro focal myocardial infarction.

Methods: Thirty patients with a diagnosis of ischemic heart diseases and myocardial infarction were selected. Fifteen of them were treated with the standard protocol of treatment (control group). The other 15 were transplanted with their bone marrow stem cells. Stem cells CD133 were isolated from mononuclear cells using Ficoll followed by immuno-magnetic separation. Isolated cells were injected in intra-arterial into the coronary arteries under angiography in the average dose of 5ml of suspension containing 0.8-1.5 million cells. Clinical examination and current tests such as ECG, LVEF, ESV, EDV were performed. In order to evaluate the dynamics of myocardial perfusion we carried out myocardial scintigraphy using Tc99m with the ligand methoxyisobutylisonitrile (MIBI).

Results: Key indicators of hemodynamic of heart were recorded before and after six months of the introduction of autologous stem cells. As presented in the table, the ejection fraction of left ventricle and end-diastolic volume of the left ventricle had a clear tendency to normalization.

Conclusions: Our method of stem cell transplantation is safe and does not increase mortality as a consequence of heart disease. Treatment of autologous stem cells significantly improved key indicators of heart hemodynamic. We thank J Al Majid from Dubai and H Sadeghi from Cardiovascular Surgery, University of Lausanne, for their precious help.

Z Rahimov1, A Barotov1, A Dustov2, M Khidirov3, T Gulmuradov1, G Mirojov2, B Karimzade2, N Navjuvonov 1, N Olimov1, E Rizoev1, A Gaibov2, M Rahmatov1, S Muminjonov2, A Didary3, J Irgashev3, A Therwath4,O Bobokhojaev1, U Kurbonov3, S Rahmonov1, M Mirshahi3,4.

1 Deptartment of Cardiology, Cardiovascular Surgery and Hematology of Ministry of Health of the Republic of Tajikistan, 2Institute of Gastroenterology of Tajikistan Academy of Medical Science, Dushanbe, Tajikistan, 3Deptartment of Surgical Diseases and Stem Cell Laboratory, Avicenna Tajik State Medical University, 4 UMRS 872, INSERM, Paris VI Faculty of Medicine, Paris, France

In the United States, nearly six million people currently suffer from heart failure. Organ transplantation remains the only definitive treatment for end-organ failure. However, the supply of donor organs is limited. Lifelong immunosuppression and chronic rejection limit widespread clinical impact of organ transplantation. The creation of an autologous bioartificial heart with preserved coronary and three-dimensional architecture could theoretically bypass these issues by providing a source of donor organs. Anterograde coronary perfusion of detergents (Sodium Dodecyl Sulfate: SDS, De-ionized water, and Triton-X) produced acellular heart scaffolds from small (rats) and human-sized (porcine and human) hearts. Perfusion-decellularized hearts were void of cellular content (DNA) and perfusing detergents (SDS) but had preserved coronary anatomy and gross structural integrity (sGAG quantification; immunohistochemistry for matrix proteins Collagen I, III, IV, X, Laminin, Fibronectin, Elastin).

Perfusion-decellularized whole heart scaffolds are biocompatible (ability to act as a substrate to support the appropriate cellular activity) when repopulated with neonatal rat cardiomyocytes1 or human induced-pluripotent stem cells (hiPSCs) as evident by cellular attachment and expansion. When seeded onto perfusion-decellularized heart scaffold slices, hiPSCs-derived cardiomyocytes form functional myocardial tissue, in vitro (visible spontaneous beating; immunofluorescence for myosin heavy chain and connexin-43).

After the first isolation of human embryonic stem cells was reported in 1998, many countries reviewed their national policies regarding human embryo research. During this process, several countries amended current policies or created new policies to specifically address human embryonic stem cell research. These policies can be classified in three broad categories: restrictive, permissive with limits, or permissive. Over this 13 year period, there has been a global trend to transition policies from restrictive to more permissive. These shifts have occurred due to the efforts of scientists who have advocated for policy harmonization. While countries still possess a range of policies, this attempt towards harmonization has had a positive impact on the field by improving the quality and quantity of international scientific collaborations. This talk will review national stem cell policies, describe how international collaborations have been fostered under such policies, and discuss the impact of these international collaborations in the field of stem cell research.

Over the past several years the funding of stem cell research has often been justified on the grounds that it could serve as an engine of regional economic growth. To some degree, this is part of a larger trend, one that has placed biomedical research as a central component of many jurisdictions’ economic strategies. This has led to increasing pressure on researchers to build links with industry, to rapidly move their work to the clinic and to develop commercializable products. While there are undeniable benefits with this approach, it has also created some interesting policy challenges. In this presentation I will both review the nature and extent of the commercialization pressure and explore the related legal, ethical and social issues, including the possible adverse impact commercialization pressure may have on the research environment and the promotion of inappropriate 'hype'.

Compelling evidence exists that somatic stem cell-based therapies protect the central nervous system (CNS) from chronic inflammation-driven degeneration, such as that occurring in experimental autoimmune encephalomyelitis (EAE), multiple sclerosis (MS) and cerebral ischemic/hemorrhagic stroke. However, while it was first assumed that stem cells may act through direct replacement of lost/damaged cells, it has now become clear that they are able to protect the damaged nervous system through a number of ‘bystander’ mechanisms other than cell replacement. In immune-mediated experimental demyelination and stroke – both in rodents and non-human primates – others and we have shown that transplanted neural stem/precursor cells (NPCs) possess a constitutive and inducible ability to mediate efficient ‘bystander’ myelin repair and axonal rescue. Yet a comprehensive understanding of the multiple mechanisms by which NPCs exert their therapeutic impact is lacking. We envisage that the remarkable therapeutic plasticity of NPCs results from their capacity to engage highly sophisticated programmes of horizontal cell-to-cell communication at the level of the (micro) environment and we attribute a key role to the transfer of secreted membrane vesicles (MVs) from (donor) NPCs to (recipient) neighbouring cells. We are starting to define whether this form of communication is biologically relevant for NPCs, and look forward to establishing whether it is associated to cell-to-cell trafficking of non-coding RNAs (ncRNAs), and indeed on elucidating its molecular signature and therapeutic significance for MS. We believe that the true innovation of this approach relies in its unique peculiarity to look into an innate cellular mechanism with the visionary focus of translating the knowledge of basal stem cell functions into innovative high-impact clinical therapeutics for MS.

Currently, there are few effective therapies for the treatment of neurodegenerative diseases or injuries to the brain, spinal cord and eye. Human neural stem cell transplants offer the prospect to treat such conditions and represent a potential exciting new medical therapy. A highly purified composition of human neural stem cells has been isolated, expanded and stored as banks of cells, HuCNS-SC®. When transplanted into the brain of immunodeficient rodents, human neural stem cells reside and proliferate in host neurogenic sites, such as the subventricular zone and dentate gyrus of the hippocampus. Their progeny migrate globally throughout the brain and differentiate in a site-appropriate manner into neurons, astrocytes and oligodendrocytes. When transplanted into the spinal cord above and below the injury site, these cells also migrate extensively and differentiate, remyelinate and make synaptic connections with host neurons. HuCNS-SC have also been shown to produce soluble proteins such as housekeeping lysosomal enzymes, neurotrophic factors, and chemokines which may protect damaged host cells and also become mature oligodendrocytes which myelinate dys- or demyelinated host axons. Moreover, these human cells survive long-term in the host brain with no signs of tumor formation or adverse effects and unlike ESC or iPSC, HuCNS-SC do not require pre-differentiation prior to transplant nor do they form teratomas. Therefore, a single transplant of human neural stem cells offers the prospect of a durable clinical benefit. Studies transplanting HuCNS-SC into animal models of human diseases or injury have been performed to assess the cells’ biological properties including their impact on these specific targets. These preclinical efficacy studies have demonstrated protection of host cells and/or improvements in specific functional deficits and provided the foundation for the neuroprotection and neural replacement strategies to support initiation of our clinical studies. Three clinical studies have been initiated to date. The first clinical study in Batten disease, a fatal lysosomal storage disease, has been completed and the surviving patients are now ~4-5 years post-transplant with no safety concerns. A trial in PMD, a fatal myelination disorder, is completing and will examine evidence of de novo myelin formation from transplanted HuCNS-SC. A trial in chronic spinal cord injury has completed dosing of the most severely injured patients and will now enrol those with incomplete thoracic injuries. The Company has recently filed regulatory documents to begin a trial in dry AMD. Preclinical studies in a rat model of retinal degeneration, the RCS rat, have shown cone photoreceptor protection following subretinal transplants of HuCNS-SC. The clinical data derived from these studies should facilitate future clinical testing of HuCNS-SC cells in a broad range of other neurological disorders including Alzheimer’s disease, stroke, and cerebral palsy. In addition to its pioneering efforts in the discovery and development of its proprietary human neural stem cell, the Company’s scientists have also identified a rare cell population from non ideal human adult livers that display the key hallmarks of a therapeutic product for treating liver diseases. Efforts are underway to define optimal expansion conditions to create cell banks of liver-engrafting cells.

Public (government) oversight reflects the way society perceives and responds to risk, which is intrinsic to the conduct of experimental research. Such oversight typically takes the form of regulation, which combines organizations, rules and sanctions intended to produce desired behaviors in support of clear objectives. Ensuring a rigorous clinical trial process that also offers proper protections of research subjects requires a legal framework that features the following qualities: (1) must act, and be seen to act, in the public interest; (2) comprised of clear and explicit objectives and requirements; (3) fairness in its application and enforcement; (4) based on dialogue among regulators, the regulated community, and the beneficiaries of regulation; (5) sustains public confidence and trust; and (6) is more likely than other options to achieve its goals. While these features will be applied to stem cell clinical trials, they are also relevant in other areas where science and technology are perceived as posing risks to individuals and/or the larger society.

Patients, scientists, regulators, and the public may each view early stage stem cell-based clinical trials from varying perspectives and interests. In this talk I explain these various points of view, their differences, and their importance to the ethics of stem cell-based clinical trials. Based on this multiple perspectives analysis, I conclude with some ethical recommendations for the conduct of early stage human trials necessary to advance stem cell science.

Stem cell research by Weill Cornell Medical College in Qatar (WCMC-Q) is subject to the ethical standards of both the Qatar Supreme Council of Health (SCH) and the National Academies of Sciences (NAS) in the United States (US). The authors examined their personal experiences with the policy-making process in Qatar and the ethical review process in both Qatar and the US. The objectives of this examination were to characterize different approaches to ethical oversight of stem cell research; consider factors underlying the different approaches; identify advantages and shortcomings of the approaches;and suggest areas where the approaches could enrich each other.

The authors identified two different approaches. First, Top-down, in which a centralized agency passes a set of regulations, often drawn from different international experiences and intended to be complete and stable. This is the model followed in Qatar. Secondly, Bottom-up, in which various agents (researchers, universities, funders, and others) reach a level of consensus and produce non-mandatory guidelines including a set of principles and accompanying specific points. Institutions later adopt the guidelines. This is the approach of the NAS.

Each approach results from the interplay of several factors, including Local experience, Organization of the state, and Political culture.

Top-down is rapidly available and provides a clear, monolithic framework, generally with few internal contradictions.

However, speed and consistency may limit flexibility. There may not be room for ethically acceptable alternative solutions if the framework poses scientific or practical difficulties. Top-down regulation runs the risk of rapid obsolescence in a dynamic field.

Bottom-up, being the product of deliberation, tends to be more flexible. However, this approach risks being hijacked by interminable discussions. Guidelines are not always immediately available. In a decentralized environment, competing guidelines may emerge. Approaches can enrich each other by establishingbasic guiding principles to inform detailed rules; avoiding too much detail that will be difficult to change; maintaining a mix of guidelines and regulation to provide certainty and flexibility while avoiding arbitrariness; adjusting according to experience of and feedback from stakeholders; and accepting imperfection in details and implementation.

The objective was to compare the religious doctrine on human stem cell research in the faiths of: Christianity, Islam, Judaism, Hinduism, and Buddhism. The method used was the analysis of theology texts. Results showed that Locus = human potentiality, sanctity of life, soul and conceptus, subject vulnerability, cell line ownership, eugenics. We concluded that an international co-operative agreement is difficult to achieve.

Over the past few years, GCC countries have witnessed remarkable social and economic growth, which is best reflected in their well-organized and efficient health care systems. With these achievements, a shift in the pattern of disease has become evident. There has been a change in a disease pattern with a falling incidence of communicable diseases, decrease of mortality and morbidity rates of infants and children and a rise in noncommunicable conditions.

Families in GCC states can benefit from new genetic approaches, but these need to be designed to conform with the community beliefs and culture. Presently, GCC communities need help in coping with the changes and challenges that genetic knowledge brings to their lives.

Understanding social beliefs, respect of the traditions set up in an Islamic community, and understanding the psychological difficulties faced by families are essential for acceptance of new technological advances. The difficulties faced by families and proposed solutions are discussed.

Ovarian cancer is a highly metastatic disease. Epithelial ovarian cancer is the most lethal gynecologic malignancy with the majority of cases being diagnosed after the disease has become metastatic. Recent reports have shown that 25% of cancerous cells within tumors have features of cancer stem cells (CSCs). CSCs have been identified on the basis of their ability for self-renewal and to have the capacity to differentiate into cancer cells and also form tumors in animal models. EMT describes a mechanism by which cells lose their epithelial characteristics and acquire more migratory mesenchymal properties. It also seems to have a key role in the acquisition of invasive and migratory properties in many types of carcinoma cells. The aim of this study is to understand the molecular changes and signaling pathways associated with ovarian cancer cell transformation which may lead to the identification of targets for novel therapeutic interventions.

OVCAR-3 and hospicells were cultured in vitro according to standard procedure. OVCAR cells were stained using anti CD133 –prominin-1 and anti CD117-APC and sorted by FACS system as positive and negative. The supernatant of OVCAR CD117+ and CD117- were analyzed by cytokine array And 174 different membrane coupled anti-cytokines along with appropriate controls were studied. The gene expression patterns of OVCAR-3 cultured with supernatant of Hospicells (clone M16) or control cells were analysed and compared by a two-colour topic-defined microarray. The products of microarray consisted of genes related to cytokines, interleukins and growth factors. The up regulated 86 codes genes were analysed by DAVID functional annotation tool.

We demonstrated that OVCAR-3 cell line for several reasons did not appear to be monoclonal. It contains tow subpopulations: one expressing CD117 and the other not. The fact that OVCAR-3 expresses at the same time CD133 and CD117 is indicative of the stem cell nature of this cell line. The cytokines secreted by CD117+ and CD117- were not the same and indicate the different profile of these cells in cell behavior. Analysis was of OVCAR gene array after 8h incubation of Hospicells by DAVID functional annotation tool intricate gpl 130 (IL6 ST) cytokine receptor interaction (IL6, IL11, OSM, LIF, CNTF ligands) and angiogenic factors expression.

We found that OVCAR-3 is apparently a suitable model for studying ovarian cancer stem cells and Epithelial to Mesenchymal Transition in Ovarian Cancer Cells.

Neural stem cells (NSC) with self-renewal and multipotent properties serve as an ideal cell source for transplantation to treat neurodegenerative insults such as Parkinson's disease. We used Agilent's and Illumina Whole Human Genome Oligonucleotide Microarray to compare the genomic profiles of human embryonic NSC at a single time point in culture, and a multicellular tissue from postmortem adult substantia nigra (SN) which are rich in dopaminergic (DA) neurons. We identified 13525 up-regulated genes in both cell types of which 3737 (27.6%) genes were up-regulated in the hENSC, 4116 (30.4%) genes were up-regulated in the human substantia nigra dopaminergic cells, and 5672 (41.93%) were significantly up-regulated in both cell population. Careful analysis of the data that emerged using DAVID has permitted us to distinguish several genes and pathways that are involved in dopaminergic (DA) differentiation, and to identify the crucial signaling pathways that direct the process of differentiation.

The set of genes expressed more highly at hENSC is enriched in molecules known or predicted to be involved in the M phase of the mitotic cell cycle. On the other hand, the genes enriched in SN cells include a different set of functional categories, namely synaptic transmission, central nervous system development, structural constituents of the myelin sheath, the internode region of axons, myelination, cell projection, cell somata, ion transport, and the voltage-gated ion channel complex. Our results were also compared with data from various databases, and between different types of arrays, Agilent versus Illumina. This approach has allowed us to confirm the consistency of our obtained results for a large number of genes that delineate the phenotypical differences of embryonic NSCs, and SN cells.

Banking of biomedical tissues or information related to them has been conducted around the world for clinical, research and/or commercial purposes. In addition to stem cells in the differentiated tissues such as cord blood and bone marrow, pluripotent stem cells are also subjects of banking. Human embryonic stem cells (hES cells) are one type of these cells, but induced pluripotent stem cells (iPS cells) are now also subjects of banking for various purposes.

In Japan, the Center for iPS Cell Research and Application (CiRA), led by Dr Shinya Yamanaka, was established in 2010 to conduct research on iPS cells. In order to realize iPS cell based therapies, CiRA is now working to establish a new iPS cell bank. The bank aims to establish clinical-grade, HLA-matched iPS cell lines from healthy volunteers and store them to provide for therapeutic or research use. Although some iPS cell banks in Japan or in other countries, including the one in CiRA, are already established, these banks store and provide basically disease-specific iPS cell lines. Therefore, the planned iPS cell bank at CiRA is unique in that it is going to store and provide clinical-grade iPS cells derived from healthy volunteers.

We have been collaborating with researchers at CiRA to establish appropriate procedures for the recruitment of healthy volunteers. We have found that there are a variety of ethical, legal, and social issues (ELSI) that need to be addressed. Since iPS cells are pluripotent stem cells, some ELSI are similar to those already identified in hES cell banks. However, there are other issues that are relatively novel and specific to iPS cells, especially when the donor is a healthy volunteer, not a patient. These ELSI identified arise because: 1. these cells are derived from healthy volunteers; 2. these cells expand infinitely; and 3. the bank deals not only with information such as genome sequence but also with other aspects of the cells.

In this presentation, we will summarize and discuss the ELSI we encountered while establishing the process of healthy-volunteer recruitment for the iPS cell bank.

It has been shown that shear stress plays a critical role in promoting endothelial cell (EC) differentiation from embryonic stem cell (ESC)-derived ECs. However, the underlying mechanisms mediating shear stress effects in this process have yet to be investigated. It has been reported that the glycocalyx component heparan sulfate proteoglycan (HSPG) mediates shear stress mechanotransduction in mature EC.

In this study, we investigated whether cell surface HSPG plays a role in shear stress modulation of EC phenotype. ESC-derived EC were subjected to shear stress (5 dyn/cm(2) ) for 8h with or without heparinase III (Hep III) that digests heparan sulfate. Immunostaining showed that ESC-derived EC surfaces contain abundant HSPG, which could be cleaved by Hep III. We observed that shear stress significantly increased the expression of vascular EC-specific marker genes (vWF, VE-cadherin, PECAM-1. The effect of shear stress on expression of tight junction protein genes (ZO-1, OCLD, CLD5) was also evaluated. Shear stress increased the expression of ZO-1 and CLD5, while it did not alter the expression of OCLD. Shear stress increased expression of vasodilatory genes (eNOS, COX-2), while it decreased the expression of the vasoconstrictive gene ET1. After reduction of HSPG with Hep III, the shear stress-induced expression of vWF, VE-cadherin, ZO-1, eNOS, and COX-2, were abolished, suggesting that shear stress-induced expression of these genes depends on HSPG. These findings indicate for the first time that HSPG is a mechanosensor mediating shear stress-induced EC differentiation from ESC-derived EC cells.