Philip Schwartz - US grants

DESCRIPTION (provided by applicant): Human embryonic stem cells can be proliferated indefinitely while retaining a normal karyotype, and can be used in vitro to study the earliest stages of differentiation. While in vitro studies (drug testing, target identification) may ultimately be medically more valuable than in vivo applications, it is the latter that has captured the imagination of US government leaders. The decision to fund research on human embryonic stem cells opens up unprecedented opportunities. The public funding takes the research out of the private sector, where it has been motivated solely by economic necessities, into the public sector, where basic research can be performed without the driving force of the marketplace. The course proposed in this application, "Human Embryonic Stem Cell Culture Training Course", will provide hands-on training for investigators to learn how to culture, manipulate, and differentiate ES cells from humans in vitro. This course will bring together some of the leading experts on ES cell technology and through comparative approaches, effectively train students in the successful culture, maintenance and manipulation of ES cells. A major long-term goal for this course will be to share and improve standard protocols. Specifically, this course will cover: 1) Demonstration of proper growth conditions for cells; 2) Proper freeze-thaw cycling and preparation of cell passages; 3) Use of co-culture techniques; 4) Use of cell separation procedures; 5) Review of basic good laboratory practices for use of human biological materials; 6) Development of protocols that support the characterization of embryonic stem cells; 7) Application of standard research protocols for directed differentiation of embryonic stem cells; 8) Employment of standard detection methods for infectious organisms or other contaminants; and 9) Cross-training of techniques used in other applications of stem cell biology. These aims will be accomplished by offering intensive 10-day courses to 12 participants chosen for their outstanding research potential. The Courses will be held semi-annually at the National Human Neural Stem Cell Resource of the Neuroscience Laboratories at the Children's Hospital of Orange County Research Institute.

[unreadable] DESCRIPTION (provided by applicant): Human embryonic stem cells (hESCs) can be proliferated indefinitely while retaining a normal karyotype, and can be used in vitro to study the earliest stages of differentiation. While in vitro studies (drug testing, target identification) may ultimately be medically more valuable than in vivo applications, it is the latter that has captured our imagination. The decision to fund research on hESCs opens up unprecedented opportunities. Public funding takes the research out of the private sector, where it has been motivated solely by economic necessities, into the public sector, where it can be performed without the driving force of the marketplace. The course proposed in this application, "CHOC Human Embryonic Stem Cell Culture Training Course", is a continuation of our current successful course which provides hands-on training for investigators to learn how to culture, manipulate, and differentiate hESCs in vitro. This course brings together some of the leading experts on hESC technology and through comparative approaches, effectively trains students in the successful culture, maintenance and manipulation of hESCs using lines WA01 and WA09 from the NIH hESC Registry. A major long term goal for this course has been to share and improve standard protocols. Specifically, this course aims are: 1) Demonstration of proper growth conditions for cells; 2) Proper freeze-thaw cycling and preparation of cell passages; 3) Use of co-culture techniques; 4) Use of cell separation procedures; 5) Review of basic good laboratory practices for use of human biological materials; 6) Development of protocols that support the characterization of embryonic stem cells; 7) Application of standard research protocols for directed differentiation of embryonic stem cells; 8) Employment of standard detection methods for infectious organisms or other contaminants; and 9) Cross-training of techniques used in other applications of stem cell biology. These aims are accomplished by offering intensive 10-day "immersion" courses to 12 participants chosen for their outstanding research potential and commitment to institute hESC research in their own laboratories. In addition, the future courses will each conclude with a day-long symposium, "Cellular Therapies in Pediatric Diseases", designed to highlight the clinical goals and obstacles for hESC research. This symposium, which focuses primarily on pediatric diseases, will be open to the general public as well as biomedical researchers and clinicians and, for the latter, will provide Continuing Medical Education credits. The courses are held annually in the Spring at the National Human Neural Stem Cell Resource of the Neuroscience Laboratories at the Children's Hospital of Orange County Research Institute. [unreadable] [unreadable] [unreadable]

DESCRIPTION (provided by applicant): The autism spectrum disorders (ASDs) comprise a set of neurodevelopmental disorders that are, at best, still poorly understood and, yet, they are the fastest growing developmental disorder in the United States. Analysis of the post-mortem brain has provided some of the most valuable data for advancing our understanding of ASD pathophysiology. In addition, neural stem cells (NSCs) can be harvested from the post- mortem ASD brain, providing a critical tool for the study of ASDs, as detailed examination of the proliferation and differentiation of NSCs derived from ASD brains, compared with those derived from normal brains, is likely to yield important data regarding the etiology of the disease and provide new avenues for therapeutic approaches. The procurement of post-mortem ASD brains, however, has proven to be difficult. Thus, the generation of sufficient numbers of ASD NSC lines has been slow. Recent advances in stem cell research now allow for this difficulty to be overcome. It is now possible to "de-differentiate" or re-program human fibroblasts to an induced pluripotent stem cell (iPSC) state;that is, a line of stem cells that can differentiate into virtually any tissue cell type, such as brain, can be created from a fibroblast cell. This raises the potential of applying this technology to fibroblasts derived from ASD patients to allow study of resulting NSCs. Since fibroblasts are readily procured, sufficient statistical power for a whole host of studies can be achieved. We will apply the reprogramming technique to both fibroblasts derived from well- characterized patients with autism as well as those derived from normal individuals. Using the recently described gene candidates for reprogramming cells, we will transduce fibroblasts using published methods and characterize their conversion to iPSCs. We will use existing cell lines or cells lines recruited with our collaborators, procured under our stem cell-specific consent, for both the re-programming methodology and the compare and contrast evaluation of resulting cell lines. Specific methodology evaluation will be accomplished using fibroblasts and NSCs, already in our repository, that were derived from the same patients. The implication for the study of the ASDs simply cannot be overstated. Efficient generation of autism NSC lines will allow for studies that have never before been possible. These include (1) studies examining the detailed pathophysiology of the autism neuron, (2) studies of sufficient statistical power that can compare and contrast the effects of autism on neuronal differentiation, and (3) studies of the influence of environmental factors on these processes. Importantly, all data generated will be made available. The lines themselves will also be made available to the scientific community through our existing stem cell repository. This will leverage our efforts for maximal benefit to patients and families affected by autism. PUBLIC HEALTH RELEVANCE: The overall purpose of this project is to overcome a critical barrier in the field of autism research;namely, accessibility to statistically relevant numbers of patient-derived neural tissues. Our novel strategy, which is based on our considerable human neural stem cell (NSC), human embryonic stem cell (ESC), and biorepository experience, is to obtain fibroblasts from specific autism and control patients, derive induced pluripotent stem cells (iPSCs) from the fibroblasts, differentiate NSCs from the iPSCs, characterize and contrast and compare these cells using gene microarray and bioinformatics techniques, and provide the cells to specific autism researchers.