Year 4

Sanford-Burnham Medical Research Institute (SBRMI) Stem Cell Research Center (SCRC) is comprised of 4 integrated sub-cores: (1) Human Stem Cell Generation, Culture, Manipulation, & Supply; (2) Human Stem Cell Characterization; (3) High-Throughput (HT) Technologies (including such advanced & automated image analysis as high-content screening); & (4) Human Stem Cell Data-Sharing, Training, & Education. The Viral Vector Core is also housed in the SCRC.
Two years ago, the Human Stem Cell Generation Sub-Core set its goal to become the largest bank of well-characterized patient starting cells & disease-modeling hiPSCs in academia. To this end, SCC has been in the midst of establishing relationships with clinical entities in San Diego (UCSD, VA Hospital, Rady Children’s Hospital) to create a pipeline for somatic cell samples. In addition, SCRC has formed relationships with investigators abroad who have banks of cells from well-characterized patients. Furthermore, to help validate potential pathophysiological pathways & novel drug targets, SCRC has partnered with large hospital-based banks of well-archived clinical specimens from patients with similar disorders. This sub-core also established the integration free reprogramming methods on human skin fibroblasts. Recently, the sub-core successfully reprogrammed immortalized B lymphocytes. This achievement is significant because (1) reprogramming immortalized B lymphocyte is considered a very difficult process (2) a large number of the patient biopsies particularly for the mental illness are preserved in the format of EBV immortalized B lymphocyte. This will enable the researchers to generate relevant disease models from these samples, which was impossible in the past. The Sub-core also formed collaborations with Intrexon to develop a high throughput reprogramming method using a computer controlled laser system.
The Characterization Sub-Core profiles hESCs, hIPSCs, & somatic stem cells using of immunocytochemistry &cytometry; gene microarrays & deep sequencing; quantitative PCR, methylation, SNP, & microRNA/SiRNA screens; large-scale comprehensive (phospho)proteomics, metabolomics, &pharmacogenetics; & high-resolution cellular imaging – all with an eye to identifying biomarkers for the diagnosis, prognosis, progression, & therapeutic options for such disorders. Via the SCRC website, the largest comparative phosphoproteomic dataset in any biological system, comparing human pluripotent stem cells with their pure neurectodermal derivatives can accessed publicly. Advanced gene microarrays, quantitative PCR, microRNA screens, & proteomics were used to provide genomic fingerprinting of an array of human stem cell lines & their differentiated progeny. “PluriTest”, a web-based method for determining whether a given cell with a particular profile of gene expression does or does not comport to being “pluripotent”, a tool that is coming to replace the use of teratoma formation as a proof of pluripotence.
One of the principal achievements of the HT Technologies Sub-Core was creating read-outs for its chemical, cDNA, or shRNA/siRNA screens based on automated HT high-content microscopy used in conjunction with the SBMRI’s robotic liquid handling systems in the chemical library screening facility of the Prebys Center for Chemical Genomics (PCCG). Such efforts have been enhanced by NIH funding of a Comprehensive Screening Center at SBMRI as well as a recent partnership with Johnson & Johnson.
Being a training facility, SCRC also provides various hands-on training courses on hESC/hiPSC maintenance, hiPSC generation, lineage-specific differentiation, etc. Newly updated protocols for stem cell differentiation & somatic cell reprogramming have been posted to the SCRC website. More recently, a Standard Operating Procude (SOP) booklet covering almost all the experiments related to the stem cell culture, manipulation, and characterization was published on the SCRC website for the training course. Currently, SCRC hosts the monthly Southern California Stem Cell Consortium, a monthly stem cell User’s meeting; a monthly Journal Club – all designed to engage researchers and & trainees throughout the region. SCRC conducts numerous tours to non-stem cell scientists, non-scientists, patient advocates, journalists, donors, high school students, & residents from the local communities who are interested in learning about stem cell biology.