Year 1

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 that are being increasingly used. In addition, 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, 57 students and postdoctoral fellows from the laboratories of 11 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. As one example, the Robey and Winoto labs are developing and optimizing in vitro culture systems to induce T cell development from human embryonic stem cells and iPS cell lines. An important parameter is the choice of the starting cell line, and a number of non-registry human ES cells lines are being compared. Therefore the availability of lab space that is separate from NIH funded labs is key to being able to carry out these studies. As another example, the Healy lab is developing new synthetic substrates for culturing human embryonic stem cells under defined conditions. They have made significant progress with several surface formulations, and quantitative measurements on the maintenance of hESC proliferation and pluripotency are being aided by the imaging equipment in the CIRM facility. As another 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. As an additional example, the Li lab has been utilizing micropatterned surfaces of variable mechanical properties to study the effects of substrate mechanics on stem cell function, and these efforts have been enabled by the high throughput imager. 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 thus anticipate that this valuable facility will thus be increasingly utilized to advance a growing number of projects.