The central mission of our Berkeley CIRM Shared Stem Cell Facility (SSCF) is to provide our East Bay users with knowledge, expertise, training, and equipment to advance scientific knowledge of human embryonic and other stem cells. Our facility is designed to support human embryonic stem cell (hESC) culture, including high quality cell culture space equipped with biosafety cabinets, incubators, cryogenic storage, and a standard microscope. In addition, we have developed and focused state-of-the-art resources and expertise to meet a growing need for our users, imaging. In particular, we have a two photon and visible confocal fluorescence microscope and a high throughput fluorescence imager (ImageXpress) that are being increasingly used. Our analytical and sorting flow cytometry capabilities complement the imaging equipment by providing high throughput cell fluorescence measurements. Finally, the facility is equipped with a laminar flow hood to conduct chemistry to create biomaterials and micropatterned surfaces for stem cell culture, as well as subsequent analysis by imaging and flow. We are very thankful to CIRM for enabling the construction and development of this state-of-the-art stem cell facility. Also, the generous resources of CIRM have been additionally leveraged, as the facility director has obtained additional campus funds to purchase additional equipment and further enhance the capabilities of the existing equipment.
Our facility is directed by Prof. David Schaffer in collaboration with the management oversight committee. In addition, it is managed by Dr. Mary West, who has successfully overseen the installation of the equipment and resources described above, provides rigorous training to our users, and aids in the development of imaging strategies to enable numerous experiments.
In greater scientific detail, 75 students and postdoctoral fellows from the laboratories of 21 PIs have approved access and are using the facility to date. Therefore, these resources are enabling and enhancing a large and growing number of research projects, which are described in greater detail in the full scientific progress report. The goal of a project in the Song Li lab that has made great progress this year is to derive functional neural crest stem cells (NCSCs) from embryonic stem cells and induced pluripotent stem cells. The derived NCSCs were used in nerve and blood vessel regeneration. In addition, other projects aimed at identification and characterization of adult stem cells in blood vessels. The marker expression and multipotency were determined by using ImageXpress and flow cytometry at CIRM SSCF. The next phase of this study focuses on the function of vascular stem cells in other diseases including arthrosclerosis. As another example, the Healy lab is developing new synthetic substrates for culturing human embryonic stem cells under defined conditions. Progress was made and published by the Healy lab in creating synthetic culture surfaces and materials for long-term hESC propagation, a project that has involved heavy usage of the ImageXpress. As a final example, the Schaffer lab has engineered new gene delivery vehicles that can augment and edit the genome of hESCs and neural stem cells. Quantification of gene delivery efficiency and gene targeting has been greatly aided with the flow cytometry and high throughput imaging capabilities of the CIRM facility. Finally, the Kumar lab studies how the molecular and mechanical properties of the cellular cytoskeleton regulate cell function. For example, he uses laser ablation of subcellular features such as cytoskeletal filaments to provide real time information on the mechanical properties of the cytoskeleton. He is establishing laser ablation using the multiphoton microscope in the CIRM facility, a capability that will be applied to study cytoskeletal mechanics of stem cells.
We anticipate in the upcoming year that this valuable facility will thus be increasingly utilized to advance a growing number of projects.