Feeder- and serum-free establishment and expansion of human induced pluripotent stem cells

Mehdi Totonchi, Adeleh Taei, Ali Seifinejad, Mohammadsharif Tabebordbar, Hassan Rassouli, Ali Farrokhi, Hamid Gourabi, Nasser Aghdami, Ghasem Hosseini-Salekdeh, Hossein Baharvand*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

68 Citations (Scopus)

Abstract

Although human induced pluripotent stem cells (hiPSCs) hold great promise as a source of differentiated cells for vast therapeutic implications, many obstacles still need to be surmounted before this can become a reality. One obstacle, a robust feeder- and serum-free system to generate and expand hiPSCs in culture is still unavailable. Here, for the first time, we describe a novel establishment and maintenance culture technique that uses human dermal fibroblasts to generate hiPSCs by introducing four factors, Klf4, Oct4, Sox2, and c-Myc under serum- and feeder-independent conditions. We have used a serum replacement product, conditioned medium (CM), or feeder-free medium (FFM) supplemented with high elevated basic-fibroblast growth factor in the absence or presence of Matrigel. Our FFM system in the presence of Matrigel enhanced the efficiency of alkaline phosphatase-positive colonies at a frequency at least 10-fold greater than the conventional method on feeder cells. The established hiPSCs are similar to human embryonic stem cells in many aspects including morphology, passaging, surface and pluripotency markers, normal karyotype, gene expression, ultrastructure, and in vitro differentiation. Such hiPSCs could be useful particularly in the context of in vitro disease modeling, pharmaceutical screening and in cellular replacement therapies once the safety issues have been overcome.

Original languageEnglish
Pages (from-to)877-886
Number of pages10
JournalInternational Journal of Developmental Biology
Volume54
Issue number5
DOIs
Publication statusPublished - 1 Dec 2010
Externally publishedYes

Keywords

  • Derivation
  • Maintenance
  • Serum and feeder-free culture condition

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