Year 1
The UCSD Human Embryonic Stem Cell Core Facility (HESCCF, also referred to as “the Core”) provides the infrastructure for UCSD scientists to launch and expand their research projects with human pluripotent stem cells (hPSCs, which include embryonic and induced pluripotent stem cells). Over the past four years (2006-2010), the staff of the core facility has trained over 100 researchers in the basic methods of human embryonic stem cell growth and differentiation. The staff has also conducted advanced scientific projects that aim to create the specialized cells needed by researchers and investigators. The shared research lab has served as a staging area for research by over 45 labs and over 100 individual researchers. These researchers are from all areas of the university – biology, neuroscience, bioengineering, material sciences, medicine, just to name a few.
Management and use of the laboratory
The HESCCF provides biosafety cabinets, incubators, microscopes, -150°C freezers, mycoplasma testing, karyotyping, MEFs (supportive feeder cells for hPSCs), quarantine and other control practices, sharing of protocols & best practices and specialized technologies (flow cytometry, confocal microscopy, automated microscopy, electrophysiological equipment). During this reporting period, in which renovations were completed, we were able to provide all of these services and technologies without disruption, by using interim space and CIRM-supported personnel.
Renovations in CMM East were completed on September 2, 2009, and HESCCF operations returned all activities to the renovated space. To provide optimal stem cell services and innovative products, the HESCCF received a Stem Cell Program subsidy from the UCSD Stem Cell Program in addition to the CIRM operating funds received from CIRM.
This facility runs as a small non-profit business for the benefit of California researchers at UCSD and neighboring institutes. All services and technologies are available on a recharge mechanism with the longer-term goal of sustaining operations beyond the funding period of this grant.
Coming Advances
With developments in the political landscape since the Obama executive order of March 2009 and the fast pace of research and discovery, the mission of the HESCCF is shifting to enable novel technology development critical to stem cell research. Our present mission has the following three parts:
• Provide access to specialized shared equipment. The HESCCF’s high value technologies, including cell sorting and analysis, confocal microscopy and electrophysiology, enable the research of stem cell scientists at UCSD. These technologies and the cell culture, molecular, and cell biological resources of the Core are available to all users at approved rates.
• Offer a location for “fast start” and preliminary studies by newly recruited faculty and current faculty who are finding that using hESC offers advantages not met by their current methods. The Core offers expertise and courses targeting stem cell methods that allow scientists to quickly generate preliminary data essential for planning future research and obtaining funding.
• Contributing to the development of novel technologies. HESCCF resources and staff are dedicated to maintaining state-of-the-art technology and training. Services are performed and developed in close collaboration with scientists at UCSD.
Noteworthy outcomes resulting from this project.
Human Non-Embryonic Pluripotent Stem Cell Lines
Core facility staff research associates have established the basic methods for human iPSC generation. Both retro- and lenti-viral methods for gene transduction of the reprogramming factors been established, and a number of iPSC lines have been generated from human foreskin fibroblasts. With the assistance of the core facility staff, users of the core facility can quickly launch research programs in the iPSC field.
Genetically Engineered Human Stem Cells
Staff scientists of the Core have generated a human embryonic stem cell line, which permits the targeted insertion of genetic elements into a specific locus on Chromosome 13. This “entry cell line” and the technology to target this chromosomal locus with genes of interest are available to all users of the Core.
Novel Tools or Technologies
The Core has established the basic methodology for homologous recombination in hPSCs using recombinant adeno-associated viral vectors (rAAV). To date two genes have been targeted for gene replacement using this method. In collaboration with stem cell researchers, the Core will facilitate the further development of this technology and generate a panel of hPSC marker cell lines.
Outreach Activities
Members of the public visited the CIRM UCSD HESCCF during the following events:
• Sweetwater and San Dieguito High schools visit for Science on the Mesa, April 1, 2009
• NAS High School teacher Stem Cell Workshop, April 25, 2009
• Donor Event Lab tours, May 21, 2009
• Stem Cell Awareness Day, Sept 23, 2009