Induced Pluripotent Stem Cell Core (JDC)
Director: Wagers, Amy, Ph.D.
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.
Director: Wagers, Amy, Ph.D.
Phone: (617) 732-2590
Kulkarni, Rohit, Ph.D., M.D.
Role: Associate Director, Induced Pluripotent Stem Cell Core, Associate Professor of Medicine, Harvard Medical School, Investigator, Joslin Diabetes Center
Phone: (617) 713-3460
Goldfine, Allison B., M.D.
Role: Associate Director, Induced Pluripotent Stem Cell Core
- 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.
- 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."
- iPS cell line expansion, banking, and distribution
- Isolation of DNA, RNA or protein purified from iPS lines
- 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.
- Patient identification
"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."
- 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.
- 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.
- 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.
- Validation and quality control of reprogrammed iPS lines
The following characterization and quality control services can be selected.
Pluripotency verification through:
Flow Cytometry analysis
Real-Time PCR analysis
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