Induced Pluripotent Stem Cell Core
Induced pluripotent stem cells (iPS cells), generated by transcription factor-dependent nuclear reprogramming of differentiated somatic cells, are pluripotent stem cell lines that can be propagated indefinitely in culture and maintain the potential to differentiate into any cell type in the body. As iPS cells retain the same genetic make-up as the somatic cell targeted for reprogramming, these cells hold tremendous promise for uncovering novel genetic and biochemical factors that underlie diseases with complex and poorly understood genetic influences, such as diabetes. The newly established iPS Core maintains a centralized facility for the reliable and consistent generation and propagation of reprogrammed iPS cells for use in cutting-edge research into the molecular and cellular pathologies underlying diabetes and its complications.
1. Experimental design and IRB documentation. New and existing DRC investigators who wish to employ iPS cells in their experiments should begin by contacting the iPS Core for consultation. An initial meeting will be scheduled involving the Core Director, Associate Directors, the Core’s Clinical Research Nurse and the investigator, in which the aims, experimental design, approach, and requirements for human subjects research are discussed.
2. Patients. Assistance in identifying patients who fit study criteria for the specific aims of the investigators using the Core from among Joslin’s patient population and through local advertising (flyers in Joslin public areas, web-postings, etc.) using IRB-approved postings.
3. Patient Samples. Skin punch biopsies, peripheral blood, or other pertinent tissue will be obtained under informed consent for reprogramming studies. Punch biopsy and/or venapuncture will be performed in Joslin’s Clinical laboratory (located on the 3rd floor of the Joslin Diabetes Center), by trained medical staff.
4. Sample cell culture, expansion and storage. Fibroblasts will be cultured from human skin biopsies under standard conditions. Mononuclear cells from peripheral blood can be expanded by a variety of methods. Researchers who wish to generate iPSC lines from blood samples should consult with lab personnel to determine the optimal cell population for expansion and reprogramming that will best suit their needs. All samples collected and expanded will be cryopreserved and banked at Joslin.
5. Non-integrative Reprogramming. Human iPSC lines will be generated from cultured fibroblasts, peripheral blood, or other relevant cell types using Sendai virus or episomal plasmid transfection.
6.Validation and quality control of iPS lines. The following characterization and quality control services can be selected:
- Pluripotency verification through:
- Flow Cytometry analysis
- Immunofluorescence imaging
- Real-Time PCR analysis
- Teratoma formation
- Mycoplasma testing
7. iPS cell line expansion, banking, and distribution.
8. Isolation of DNA, RNA or protein purified from iPS lines.
9. Discounted iPS culture reagents and media preparation. Joslin users can purchase discounted iPS culture reagents from a variety of vendors directly through Joslin's Cortex Website. The core can also prepare culture media as a for-fee service.
10. Training in iPS cell culture and subsequent use of core facilities for self run studies. Workshops for iPS culture training are scheduled bi-annually. For information on protocols and techniques used by the core, please visit http://stembook.org/protocols/pluripotent-cells.
The Joslin iPS core is a member of the COREdinates stem cell consortium, a group of stem cell cores working to standardize human pluripotent stem cell practices to facilitate research. If you are interested in learning more about this consortium please visit our website at http://coredinates.org/index.html.
For questions about the iPS Core and its services, please contact Amy Wagers: Amy.Wagers@joslin.harvard.edu or email: iPSCore@joslin.harvard.edu.
Page last updated: February 27, 2015