|Su-Chun Zhang, MD, PhD
iPSC Service Director
|Anita Bhattacharyya, PhD
iPSC Service Co-Director
iPSC Service Staff
iPSC Service Staff
Cell reprogramming from biopsy tissue, banked or fresh fibroblasts will be carried out by the induced Pluripotent Stem Cell (iPSC) service of the Waisman Center Cellular and Molecular Neuroscience (WC CMN) Core under the direction of Dr. Su-Chun Zhang1 . Since 2009, the WC CMN Core iPSC service has successfully reprogrammed over 30 samples into iPSCs.
All projects must obtain prior institutional IRB approval (human subjects, biosafety, stem cells, if applicable). In addition, funding information must be provided prior to initiation of reprogramming. Upon receipt, the starting cell sample will be expanded in quarantine until confirmed negative for mycoplasma infection. We recommend that the starting cell sample be karyotyped early during the expansion phase to serve as a reference point for characterization of the reprogrammed clones2. Initial reprogramming involves the electroporation of fibroblasts with episomal plasmids containing genes for the transcription factors Oct4, Sox2, Klf4 and Myc3. Transfected fibroblasts are then cultured for approximately 3 weeks prior to selection of iPSC colonies. Up to 3 individual iPSC clones are selected for characterization and are passaged until stable lines are established. Once established, iPSC clones are karyotyped2 and compared to the initial fibroblast sample. Selected clones are also analyzed for pluripotency marker expression by immunofluorescence (Sox-2, Oct-4, NANOG, Tra-1-81)4. All clones will be frozen, test thawed and tested for mycoplasma. After characterization is completed, all clones and results will be transferred to the investigator and the project will be billed.
1 This service is supported in part by a core grant to the Waisman Center from the National Institute of Child Health and Human Development (P30 HD03352).
2 The iPSC service employs WiCell Cytogenetics for all karyotyping (separate charges apply).
3 Okita et al., Nature Methods. 2011 May;8(5):409-12. Reprogramming and culture methods can be adjusted to investigator’s needs.
4 Additional pluripotency testing by flow cytometry is available upon request (separate charges apply).
iPSC Service TIMELINE
|Generation of fibroblast cell lines from researcher-provided materials||2-6 weeks|
|Reprogramming of cells||3-6 weeks|
|Selection and expansion of iPSC clones, freeze, and characterization||2-6 weeks|
Due to the experimental nature of the materials provided to the Waisman Center by the researchers, delays in performing the reprogramming services or the inability to complete the services, which are not unreasonable under comparable industry standards, shall not be the cause of any claim against the Waisman Center.
CONTACT: Email Anita Bhattacharyya, Ph.D. or call 608.265.6142