Summary of NIH Stem Cell Unit 4/11/2005 Meeting
NIH Stem Cell Characterization Unit
Steering Committee Conference Call
Monday, April 11, 2005
10:00 a.m. EST
Participants: Allen M. Spiegel, M.D. (co-chair, NIH Stem Cell Task Force); Ronald D.G. McKay, Ph.D. (Unit Director); James F. Battey, Jr., M.D., Ph.D.; David M. Bodine, Ph.D.; Curt Civin, M.D.; Marvin C. Gershengorn, M.D.; Robert G. Hawley, Ph.D.; Brigid L.M. Hogan, Ph.D.; Paul Meltzer, M.D., Ph.D.; Mahendra Rao, M.D., Ph.D.; Janet Rossant, Ph.D.; Jonathan Vogel, M.D.
Unable to Participate: Elizabeth Nabel, M.D.; Story C. Landis, Ph.D.
- Welcome and Introductions—Dr. Battey, Director, NIDCD
Dr. Battey opened the meeting by thanking everyone for taking the time to help oversee the operation of the NIH Stem Cell Characterization Unit. He also thanked Dr. Paul Meltzer of NHGRI for serving as an ad-hoc advisor for this meeting. Participants introduced themselves.
- Ongoing Efforts to Monitor the Stability of the Human Embryonic Stem Cell Lines on the NIH Stem Cell Registry
- Dr. McKay gave a summary of the Unit's work thus far to characterize the Registry lines. He described three phases of their work. In phase one, the Unit acquired the hESC lines, cultured them using the provider's protocol, established frozen stocks, and then developed a standard protocol for culturing all of the cell lines. Lines were subjected to karyotype and FACS analysis. In phase two, the Unit scientists developed subcloning techniques and analyzed the hESCs using high resolution genome scanning. They contracted with the Cold Spring Harbor group to use their ROMA (Representational Oligonucleotide Microarray Analysis) technique (see Lucito et al. Genome Research 13:2291–2305). Now in phase three, the group is collaborating with other stem cell research groups worldwide with the goal of producing a "snapshot" of cells grown around the world. The Unit has shared data on 4 cell lines with the international stem cell effort being led by the United Kingdom (UK). The UK has provided standardized antibodies for use in FACS analysis of hESC surface antigens. Within the next 4 months, the international effort will be able to describe the stability of the hESCs by analyzing DNA from embryoid bodies and undifferentiated cells (all grown using feeder cells) using a 50-gene microarray. They hope to report these data at the First International Human ES Cell Workshop to be held at The Jackson Laboratory from Aug. 4–7, 2005.
- Dr. Rao summarized his group's hESC characterization efforts, which complement those ongoing in the NIH Stem Cell Unit. They used matched pairs of early and late passage hESC DNA samples to compare sequence of mitochondrial DNA, methylation patterns and high resolution karyotyping. A collaborator is examining X-chromosome inactivation in early vs. late-passage hESCs.
They conclude that although the embryonic stem cells are much more stable than other human stem cell populations, ongoing routine DNA testing is needed to monitor stability.
- Discussion and Recommendations—all participants
The participants discussed pros and cons of various available technologies for monitoring the stability of the hESCs in culture. Dr. Meltzer suggested that the Unit consider using Comparative Genome Hybridization (CGH) for the assays because of ease in dissemination, reduced cost, and the commercial availability of high resolution arrays.
The group agreed that Drs. Bodine, Meltzer, McKay, and Battey will meet again to set up a pilot study to test the use of the CGH technology on the hESCs listed on the NIH Stem Cell Registry. The pilot test will involve cells grown in feeder-free conditions.
The agenda for the next meeting (date and time to be determined) will include discussion of scale-up of hESCs in culture.
Dr. Battey once again thanked the participants for their time and contributions. The conference call concluded at 10:55 a.m.