iPSC reprogramming

Learn how to reprogram induced pluripotent stem cells (iPSCs).

iPSCs can be induced by forcing somatic cells to express transcription factors associated with pluripotency eg Oct4, Klf4, Sox2, cMyc, etc (Takahashi et al., 2006). Many different methods have been successfully used to express different combinations of these pluripotency factors resulting in effectively reprogrammed iPSCs. Reprogramming of iPSCs is usually carried out by either chemical methods (Huangfu et al., 2008) or lentiviral and retroviral transfection to incorporate DNA encoding the pluripotency genes into the host genome (Yu et al., 2007).

We provide many different biochemicals you can use to reprogram somatic adult cells into iPSCs. Some key examples of this include the following:

Human breast cancer cells (MDA-MB-468) can be reprogrammed using a combination of rapamycin (mTOR inhibitor) and Y27632 (ROCK inhibitor) with 90% efficacy after seven days of induction (Yuan et al., 2018). This could be achieved using 2 μM Y27632 and rapamycin.

Glycogen synthase kinase 3 (GSK-3) inhibitor, CHIR99021, can be used to reprogram mouse embryonic fibroblasts (MEFs) already transduced to express Oct4 and Klf4 (Li et al., 2009). This was the first example of an iPSC generated without the expression of Sox2. The MEF cells we treated with 10 μM CHIR99021 for four weeks.

BIX 01294, histone methyltransferase inhibitor can be used to reprogram mouse embryonic fibroblasts (MEFs) into PSCs when combined with Oct4 and Klf4 expression (Shi et al., 2008)

References

  1. Takahashi, K.,, & Yamanaka, S. Induction of Pluripotent Stem Cells from Mouse Embryonic and Adult Fibroblast Cultures by Defined Factors. Cell 126 (4),663-676 (2006)
  2. Huangfu, D.,, Maehr, R., , Guo, W.,, et. al. Induction of pluripotent stem cells by defined factors is greatly improved by small-molecule compounds. Nature Biotechnology  26 (7),795-797 (2008)
  3. Yu, J.,, Vodyanik, M. A., , et. al, Induced pluripotent stem cell lines derived from human somatic cells. Science  3318 (5858),1917-1920 (2007)
  4. Yuan, J.,, Zhang, F.,, You, M., , & Yang, Q. Identification of protein kinase inhibitors to reprogram breast cancer cells. Cell Death and Disease  9 ,9 (2018)
  5. Li, W.,, Zhou, H., , et. al. Generation of human-induced pluripotent stem cells in the absence of exogenous Sox2. Stem Cells  27 (12),2992-3000 (2009)
  6. Shi, Y., , Desponts, C., , et. al. Induction of Pluripotent Stem Cells from Mouse Embryonic Fibroblasts by Oct4 and Klf4 with Small-Molecule Compounds. Cell Stem Cell  3 (5),568-574 (2008)