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Targeted Projects in Human Embryonic Stem Cell Research - 2009

RFA #: 0903091046


PI Amount Title
The New York Stem Cell Foundation Dieter Egli $931,586 Derivation of Pluripotent Human Stem Cells by Somatic Cell Nuclear Transfer
The New York Stem Cell Foundation Scott Noggle $980,364 Derivation of Genetically Diseased Human Embryonic Stem Cell Lines
SUNY – University at Albany Janet Paluh $992,553 Derivation of Xenofree Human Embryonic Stem Cell Lines from Minority Populations


Derivation of Pluripotent Human Stem Cells by Somatic Cell Nuclear Transfer

Dieter Egli, PhD
The New York Stem Cell Foundation

The adult body has a limited ability to regenerate lost or diseased cells. A major goal of stem cell research is to create cells that can provide that regenerative capacity. Embryonic stem cells (ESCs) may be suitable for this purpose because they have the ability to generate all cell types of the body, including pancreatic beta cells, the cells affected in diabetes. Human ESCs derived by nuclear transfer may differ from induced pluripotent stem cells (iPSCs) in ways that will favor use of somatic cell nuclear transfer (SCNT)-derived cells for cellular therapies in humans. The derivation of human ESCs by nuclear transfer has thus far not been achieved because of a lack of systematic and controlled studies on this subject. Legal restrictions, a lack in funding, and limited access to human oocytes have permitted only a small number of studies. These studies did not allow a conclusion as to why development after nuclear transfer into human oocytes has failed. We have established IRB protocols that allow us the unique opportunity to overcome these logistical challenges and perform SCNT experiments. Here we propose to 1) resolve the cause of developmental arrest after somatic cell nuclear transfer. And 2) use this information to establish an efficient method for nuclear transfer into human oocytes. We expect that this will allow us to routinely derive nuclear transfer stem cell lines. These cell lines will then be examined for their quality both molecularly and functionally. These experiments are relevant regarding the use of reprogrammed cells for cell replacement. A major challenge in translating the potential of stem cells into therapeutic applications is to control for quality to minimize risks. The establishment of a reprogramming technique by nuclear transfer will establish a quality standard for reprogrammed cells and allow a comparison with iPS cells. Only a detailed analysis regarding the quality of reprogrammed cells will inform the field on how and which cells to select for both research and cell replacement. An intended extension of this work is the use of nuclear transfer ESCs for cell replacement for patients with diabetes, illustrating the broad relevance of this proposal.

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Derivation of Genetically Diseased Human Embryonic Stem Cell Lines

Scott Noggle, PhD
The New York Stem Cell Foundation

The capacity of human embryonic stem cells (hESCs) to self-renew indefinitely in culture while retaining their ability to differentiate into all cell types in the body suggests that they have enormous utility both in medical applications and as a research tool for addressing fundamental questions in human biology and disease. hESCs differentiated in vitro into cell types lost in a particular disease might be used for cell replacement therapies or studied to allow a greater understanding of both developmental processes and disease progression. We propose to develop new methods for deriving hESC lines from embryos with genetic diseases. We have developed efficient protocols for the isolation of new hESC lines. As the limited resource of human embryos donated after preimplantation genetic diagnosis requires maximizing derivation efficiency, we aim to use these protocols to expand the number of disease specific hESC lines available for disease modeling and research. We propose to optimize protocols to derive new disease-specific hESC lines from embryos in excess of patient need. We will derive a population of hESC lines genetically diagnosed to be affected by spinal muscular atrophy (SMA) as well as cell lines carrying Huntington's Disease (HD) with different CAG repeat lengths. Once these cell lines are generated, we will test whether there are deficiencies in development or survival of affected neuronal populations in in vitro models of these diseases. In the long term, by understanding the optimal methods for derivation of hESC lines, these methods will increase the probability of successful derivation from rare embryos such as those obtained after preimplantation genetic diagnosis. This will result in a sizeable population of disease-specific cell lines available for in vitro disease modeling and drug discovery.

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Derivation of Xenofree Human Embryonic Stem Cell Lines from Minority Populations

Janet Paluh, PhD
SUNY - University at Albany

Pluripotent stem cells in human therapies offer unrivaled potential. Human embryonic stem cells (hESCs) currently provide the most reliable source of such cells although their use involves moral/ethical considerations and is complicated by immune status. Current hESC lines reflect limited ethnic diversity, particularly among minorities, and many are unable to be used in therapies due to culturing artifacts, high passage number or regulatory restrictions. The current study will generate new hESC lines from human oocytes from minority populations under xenofree conditions and current regulatory guidelines to promote their future use in clinical trials. We will generate a set of hESC lines derived under identical conditions, characterized using the same platform and at the same passage that will allow us to determine whether ethnic background has an impact on hESC characteristics. We hypothesize that hESCs derived from embryos that have different ethnic origin will have distinctive molecular features that differentiate them. New hESC lines will be derived using patient donated embryos from minority populations. Excess embryos are donated upon completion of families following in vitro fertilization. We will use xenofree conditions with current protocols and the guidance of a fertility expert on handling embryos. These lines will be made available to the general scientific community. If our hypothesis is correct, it will underscore the need for further developing ethnic-specific medical research and treatments as well as disease prevention plans. Having hESC lines available from these racial groups will facilitate the discovery of new medical treatments targeted to specific underrepresented and minority human populations. Through our work, the scientific community will have available access to stocks of cell lines at different passages, from the very early ones, as early as passage 5-6. Researchers will not only have access to hESCs of multiple ethnic backgrounds but at different passages as well. Improving the quality, number and ethnic diversity of hESC lines derived under current national regulatory guidelines will provide an extended resource of these cells for possible therapeutic applications and for basic research with ethnically diverse lines.

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