Year 2

The UCLA Broad Stem Cell Research Center (BSCRC) Shared Research Laboratories (SRL) provide 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 40 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 of each cell line are carried with 10 different unmodified hESC lines, including 14 UCLA hESC lines on the NIH and CIRM Registries, 3 genetically modified hESC lines that carry new genes relevant to the work of several laboratories, and 18 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): 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, established operating procedures for hESC culturing and distribution, derives,expands and cryopreservers sufficient quantities of mouse embryonic fibroblasts necessary for large scale hESC culturing, and distributed ~1100 hESC plates to individual researchers.

3. iPSC Derivation Lab: 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. The Derivation Lab organizes careful characterization of all generated and distributed iPSC including: karyotyping, gene expression profiling, teratoma assay, and mycoplasma testing. The Lab & Core Bank are the only UCLA facilities providing iPSC to dozens of UCLA labs yearly. The Lab developed 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. Supported by the BSCRC and a CIRM grant, the Lab recently demonstrated that xenobiotic- free iPSC can be derived from human tissue and then differentiated providing the procedures necessary to derive GMP-compliant iPSC derivatives. (See: From Skin Biopsy to Neurons through a Pluripotent Intermediate Under Good Manufacturing Practice Protocols. Karumbayaram et. al., Stem Cells Trans Med, December 7, 2011; doi:10.5966/sctm.2011-0001)

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

5. Kohn Lab: His SRL based research supports an ADA-SCID project and a CIRM funded Disease Team project on stem cell gene therapy for Sickle Cell Disease (SCD). Data generated by this lab was incorporated into a pre-IND conference with FDA in March 2011.

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: A CIRM funded Tools & Technologies project to develop 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.