Our institution is a tertiary-care academic pediatric medical center that combines care of severely ill children, research into the causes and treatments of childhood disorders, and training of the next generation of pediatric clinical physicians, nurses and allied health care professionals and biomedical scientists. A unique focus of the research in our institution is on applications to pediatric disorders such as diabetes, inherited disorders (cystic fibrosis, muscular dystrophy, sickle cell disease, etc), cancer and congenital birth defects. It is our central hypothesis that childhood disorders will be especially responsive to therapies produced by the use of stem cells; advances in the use of stem cells to treat childhood illnesses will then lead the way to treatments for the many disorders that occur later in life. For over a decade, the Stem Cell Program at our institution has been at the leading edge of translational research for cell and gene therapy and tissue engineering, with outstanding research programs in stem cells, gene therapy, developmental biology, organogenesis and transplantation immunology. Active research programs studying adult stem cells (hematopoietic, mesenchymal, pancreatic, hepatic, pulmonary, amniotic) and human and murine embryonic stem cells, interact closely with clinical Centers of Excellence in organ and hematopoietic stem cell transplantation, diabetes, cancer and blood diseases, neonatology, as well as a full array of pediatric secondary and tertiary care programs. These academically-oriented clinical programs have a long-standing tradition of inter-weaving research and clinical trials with patient care, to develop and evaluate innovative new treatments for severe pediatric and adult disorders. A Core Laboratory for studies with human embryonic stem cells (hESC) was established in 2005, using institutional funding. The hESC Core has supported initial studies and developed a formal training program in methods for the growth of hESC; 40 scientists from 5 research institutions in Southern California have attended the training course to date. However, the technical and regulatory burdens inherent in hESC research, have significantly restricted further development of individual hESC research projects within the limited existing laboratory space at our institution. Funding is thus requested to remodel and equip approximately 3000 sq ft of existing space (2500 sq ft of usable laboratory space) to create a suite of laboratories for dedicated use as an hESC Core facility alongside shared laboratory space for investigators involved in hESC research. We anticipate the laboratories and equipment established using this grant will support the research of at least 20 scientist investigators at our institution and will be also made available to researchers at nearby institutions across Southern California.
Statement of Benefit to California:
Development of methods for regenerative medicine using human embryonic stem cells (hESC) will have wide-spread applications to improve the health for millions of Californians and tens of millions of people world-wide, by providing novel, effective therapies. Regenerative medicine may provide new treatments for diseases including diabetes mellitus, Parkinson’s disease, organ failure and injuries, inherited diseases and cancer and leukemia. The major challenge facing the field of regenerative medicine is to increase knowledge of the processes by which the mature cells of tissues (pancreas, brain, bone marrow, etc.) develop from stem cells, so that clinical approaches can be developed to produce cells suitable for transplantation. It is essential to establish laboratory facilities that can be used for research on hESC in a centralized manner that complies with all California and Federal guidelines. The hESC Core Laboratory and shared facilities to be developed based on this application will provide a resource to support research in stem cells by investigators from our institution, as well as investigators from across the Southern California region.
Year 1The renovation and equipping of the 3,000sqft California Institute for Regenerative Medicine sponsored Childrens Hospital Los Angeles Shared Stem Facility was completed in September 2009. The facility has four tissue culture suites with eleven clean tissue culture work stations, and associated equipment. The facility also has a large capacity for frozen sample storage, specialized microscopes and other equipment for stat-of-the-art molecular analyses of human stem cells. Since September 2009, the core staff has calibrated and monitored the laboratory environment and instrumentation, trained research personnel in the use of the shared equipment and performed on-going maintenance of the facility. The staff has also performed one-on-one training of personnel in the generation of human iPSC, genetic modification of human pluripotent stem cells, and routine culture of human embryonic stem cells. The CHLA Stem Cell Core staff has also maintained stocks of monitored and quality tested human embryonic stem cells including weekly, bi-weekly and monthly quality control analyses on the cells. During the first year of operation, 34 quality controlled human embryonic stem cell cultures were distributed to researchers at CHLA the first year of operation. These investigators have used these cells to develop models and study a variety of diseases including pediatric cancers (neuroblastoma, Ewings-sarcoma, retinoblastoma, lymphoma) and genetic diseases such as Cystic Fibrosis. Other researchers used the stem cells to study the genes and proteins that enable stem clel growth and expansion as well as differentiation. This provision of quality controlled stocks of human stem cells to investigators has decreased the amount of time individual research laboratories spend performing routine cell culture maintenance and quality control. In turn, this will increase the rate and efficiency of stem cell research at CHLA.
Year 2The CIRM sponsored Children’s Hospital Los Angeles Shared Stem Cell Facility underwent a leadership change and new personnel were hired in January 2011. The facility was reorganized and resupplied to support an increase in research utilization within the space. The staff has been providing hPSCs to researchers interested in working the pluripotent stem cell field, as well as protocols, education, consultation, protocol development, and hPSC culture technique training for those interested in entering the field. The stem cell core has been working with biotech industry to provide educational seminars to the researchers of CHLA on applications of tools in stem cell research. Research occurring in the Shared facility has increased since January 2011. Studies using the facility now include the development of disease models for retinoblastoma and Ewings sarcoma, including the study of genes and pathways, such as in retinoblastoma tumorigenesis, sympathoadrenal differentiation, and the role of the Wnt signaling pathway in hESCs. Additional studies are being developed include a disease model of intestinal development and inflammation, and stable gene knock-downs in hPSCs. The Shared facility has also been involved in assisting with developing a potential stem cell mediated therapy.
Year 3Over the last reporting period, the Children's Hospital Los Angeles Shared Research Lab (SRL) has been involved with human pluripotent stem cell research, adult stem cell research, and mouse embryonic stem cell research involving CHLA, USC, and the City of Hope research institutions. We are also participating in collaborative research with the USC Stem Cell Core facility on projects that aims to understand the biological properties of hPSC and to develop improved technologies for working with these cells. The CHLA (SRL) provides comprehensive support for researchers in the Los Angeles area, offering services including quality controlled cells and reagents, shared equipment, laboratory space, technical assistance, consultation, biobanking and training. Although the CHLA SRL does not run a formal CIRM funded techniques course, CHLA in collaboration with Dr. Victoria Fox from the USC Stem Cell Core Facility offered a two and a half day workshop for CHLA investigators wishing to learn pluripotent stem cell culture techniques. We plan to offer this course a twice a year--open to CHLA and non-CHLA researchers. The SRL also provided significant one-on-one training to CHLA researchers and support for students attending two newly established high school research program at USC: the EiHS Summer Program in Stem Cell Research and the CIRM STAR program, which is supported by a CIRM funded creativity award. We will continue to mentor and support this student though the next academic year. The stem cell core was instrumental in mentoring and supporting CIRM bridges students during the last year. We provided support, knowledge, and education regarding their projects and technical assistance with cell expansion and biobanking.
- J Cell Sci (2012) Eya1 protein phosphatase regulates tight junction formation in lung distal epithelium. (PubMed: 22685326)
- J Histochem Cytochem (2012) Numb Expression and Asymmetric Versus Symmetric Cell Division in Distal Embryonic Lung Epithelium. (PubMed: 22713487)
- Am J Physiol Lung Cell Mol Physiol (2012) Molecular mechanisms of MMP9 overexpression and its role in emphysema pathogenesis of Smad3-deficient mice. (PubMed: 22610349)
- Regen Med (2011) VEGF optimizes the formation of tissue-engineered small intestine. (PubMed: 21916592)
- J Surg Res (2011) Mesenchymal-Specific Inhibition of Vascular Endothelial Growth Factor (VEGF) Attenuates Growth in Neonatal Mice. (PubMed: 21696760)
- Mol Ther (2010) Regulated expansion of human pancreatic beta-cells. (PubMed: 20389286)