Year 6

The UCLA Broad Stem Cell Research Center (BSCRC) Shared Research Laboratories (SRL) provide critical state-of-the-art FDA compliant GMP-GTP facilities for the experimental manipulation and clinical application of human pluripotent stem cells (hPSC). The SRL also includes shared hPSC laboratories and resources for intra and extra-mural investigators engaged in stem cell research. The BSCRC-SRL is not subject to federal hESC restrictions. The progress to date includes:

1. hPSC Core Banks: The creation of Core Banks for storage and distribution of hESC & iPSC to appropriately approved UCLA investigators. To date, more than 50 investigators have received cells from the Banks. The BSCRC negotiated an agreement with WiCell to locally distribute the H1 & H9 lines directly through this Bank with a streamlined process, substantially decreasing researcher time and expense to access these important lines. Over 100 vials are carried for each of our 20 hESC lines. The Banks carry genetically modified hESC lines that have new genes relevant to the work of several laboratories, 18 UCLA hESC lines on the NIH Stem Cell Registry, and 12 UCLA iPSC lines. The Banks work closely with the BSCRC Derivation Labs to establish improved protocols for generation of clinical-grade hPSC and with the ESCRO committee to ensure compliance with CA and other requirements.

2. hESC Expansion Laboratory (EL): Supported by the BSCRC, the Lab serves as a branch of the hPSC Core Banks and is tasked with the expansion of banked hESC for distribution to qualified and approved UCLA researchers. The hESC-EL employs research technicians, establishes operating procedures for hESC culturing and distribution, derives,expands and cryopreserves sufficient quantities of mouse embryonic fibroblasts necessary for large scale hESC culturing, and distributed ~1300 hESC plates to individual researchers in CY15.

3. iPSC Derivation Lab: The BSCRC iPSC Derivation Core supported by the BSCRC, and formerly supported by a CIRM New Cell Line award (J. Zack), obtains human biological material under IRB-ESCRO approvals for iPSC reprogramming. The iPSC are characterized, expanded, and deposited in the Core Bank for storage, expansion, and distribution. Characterization of all generated and distributed iPSC includes karyotyping, gene expression profiling, teratoma assay, and mycoplasma testing. The Lab developed and published (Stem Cells Trans Med 2011) necessary procedures to derive GMP-compliant iPSC derivatives, obtain sterile biopsies and other material that are physically and enzymatically isolated under xeno-free conditions, grown in defined media, induced to a pluripotent state with defined factors free from non-human products, expanded in defined media conditions and then differentiated with GMP-compliant growth factors.

4. Shared Resources include cell analysis (LSRII), microscopy, RTPCR, and tissue processing available to SRL users.

5. Kohn Lab: His SRL based research supports ADA-SCID and Sickle Cell Disease projects (Disease Team and Early Translational awards). Direct adjacency of the SRL to the UCLA cGMP labs allows these GTP-level studies to be translated to GMP cell processing once clinical trial activities begin. Kohn also used the SRL for pre-clinical studies of cell processing for a new trial he opened in 2013 (23 subjects accrued to date) on gene therapy for ADA-deficient SCID using a lentiviral vector.

6. Crooks Lab: The research falls into three main areas: (a) Maintenance of and experiments with hESC lines; (b) Studies to develop implanted thymic microenvironments; and (c) Umbilical cord blood processing for use in a variety of hematopoietic stem cell experiments performed by the Crooks and Kohn Labs.

7. Martín Lab: Supported by a CIRM Tools & Technology grant, Martín develops both control and disease-specific iPSC from children and adults with various diseases, such as chronic diarrheal and other gastrointestinal and hepatic disorders that are presumably inherited.

8. Ribas Lab: The research is focused on engineered immunity of cancer in collaboration with Cal Tech investigators. Data from this project continues to contribute to his CIRM Disease Team II award.