Generation of induced pluripotent stem cells by a polycistronic lentiviral vector in feeder- and serum- free defined culture

Tissue Cell. 2018 Dec:55:13-24. doi: 10.1016/j.tice.2018.09.004. Epub 2018 Sep 19.

Abstract

Induced pluripotent stem cells (iPSCs) have great potentials for regenerative medicine. However, serious concerns such as the use of the viral-mediated reprogramming strategies and exposure of iPSCs to animal products from feeder cells and serum-containing medium have restricted the application of iPSCs in the clinics. Therefore, the generation of iPSCs with minimal viral integrations and in non-animal sourced and serum-free medium is necessary. In this report, a polycistronic lentiviral vector carrying Yamanaka's factors was used to reprogram mouse fibroblasts into iPSCs in feeder- and xeno-free culture environment. The generated iPSCs exhibited morphology and self-renewal properties of embryonic stem cells (ESCs), expression of specific pluripotent markers, and potentials to differentiate into the three-major distinct specialized germ layers in vitro. The iPSCs were also shown to have the potential to differentiate into neural precursor and neurons in culture, with greater than 95% expression of nestin, Pax6 and βIII-tubulin. This body of work describes an alternative method of generating iPSCs by using polycistronic lentiviral vector that may minimize the risks associated with viral vector-mediated reprogramming and animal derived products in the culture media.

Keywords: Feeder-and serum-free; Neural precursor and neuron cells; Polycistronic lentiviral vector; iPSCs.

MeSH terms

  • Animals
  • Cell Culture Techniques*
  • Culture Media*
  • Embryonic Stem Cells / cytology*
  • Feeder Cells / cytology*
  • Genetic Vectors / metabolism
  • Induced Pluripotent Stem Cells / cytology*
  • Mice, Inbred C57BL
  • Neurons / cytology
  • Regenerative Medicine / methods

Substances

  • Culture Media