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

• Main Authors: Al Abbar, Akram Faisal Mahmaud, Nordin, Norshariza, Ghazalli, Nadiah, Abdullah, Syahrilnizam
• Format: Article
• Language: English
• Published: Elsevier 2018
• Online Access: http://psasir.upm.edu.my/id/eprint/72831/1/Generation%20of%20induced%20pluripotent%20stem%20cells%20.pdf; http://psasir.upm.edu.my/id/eprint/72831/; https://pubmed.ncbi.nlm.nih.gov/30503056/
• Institution: Universiti Putra Malaysia

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.