Journal
MOLECULAR REPRODUCTION AND DEVELOPMENT
Volume 84, Issue 4, Pages 329-339Publisher
WILEY
DOI: 10.1002/mrd.22789
Keywords
canine; feeder-independent; iPS cells; serum-free
Funding
- Japan Society for the Promotion of Science [15K14888, 14J12063, 24380172]
- Japan Science and Technology Agency Competitive Funding Programs (A-STEP) [AS262Z00]
- Grants-in-Aid for Scientific Research [15K14888, 14J12063, 16H05044, 15K07747] Funding Source: KAKEN
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Canine induced pluripotent stem cells (ciPSCs) are an attractive source for regenerative veterinary medicine, and may also serve as a disease model for human regenerative medicine. Extending the application of ciPSCs from bench to bedside, however, requires resolving many issues. We generated ciPSCs expressing doxycycline-inducible murine Oct3/4 (Pou5f1), Sox2, Klf4, and c-Myc, which were introduced using lentiviral vectors. The resultant ciPSCs required doxycycline to proliferate in the undifferentiated state. Those ciPSC colonies exhibiting basic fibroblast growth factor (bFGF)-dependent proliferation were dissociated into single cells for passaging, and were maintained on a Matrigel-coated dish without feeder cells in a serum-free medium. The established ciPSCs had the ability to differentiate into three germ layers, via formation of embryoid bodies, as well as into cells expressing the same markers as mesenchymal stem cells. These ciPSCs may thus serve as a suitable source of pluripotent stem cell lines for regenerative veterinary medicine, with fewer concerns of contamination from unknown animal components.
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