Canine induced pluripotent stem cell maintenance under feeder-free and chemically-defined conditions

Mol Reprod Dev. 2021 Jun;88(6):395-404. doi: 10.1002/mrd.23478. Epub 2021 May 19.

Abstract

Canine induced pluripotent stem cells (ciPSCs) provide a platform for regenerative veterinary medicine, disease modeling, and drug discovery. However, in the conventional method, ciPSCs are maintained using chemically-undefined media containing unknown animal components under on-murine embryonic fibroblast feeder conditions, which were reported to modify cell surface of iPSCs and increases the risk of immune rejection when the cells are transplanted into patients. Moreover, in the conventional method, ciPSCs are mechanically passaged, which requires much time and effort. Therefore, the large-scale expansion of ciPSCs is difficult, which should be resolved for using ciPSCs in clinical application and research. Here, it was shown that StemFit® AK02N and iMatrix-511 could maintain the pluripotency of ciPSCs using conventional culture method. Furthermore, it was demonstrated that the feeder-free and chemically-defined ciPSC culture systems using StemFit® AK02N and iMatrix-511 could stably maintain and allow the easy expansion of ciPSCs generated using N2B27 and StemFit® AK02N, without causing karyotype abnormalities. ciPSCs expressed several pluripotency markers and formed teratomas, including cells derived from three germ layers.

Keywords: canine; chemically-defined; feeder-free; induced pluripotent stem cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biomarkers
  • Cell Adhesion
  • Cell Culture Techniques*
  • Cell Differentiation / drug effects
  • Cell Lineage
  • Cells, Cultured
  • Clone Cells
  • Coculture Techniques
  • Culture Media / analysis
  • Culture Media / pharmacology*
  • Dogs / anatomy & histology*
  • Germ Layers / cytology
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / drug effects
  • Induced Pluripotent Stem Cells / transplantation
  • Karyotyping
  • Leukocytes, Mononuclear / cytology
  • Leukocytes, Mononuclear / drug effects
  • Primary Cell Culture / methods*
  • Recombinant Proteins / pharmacology
  • Teratoma / etiology
  • Teratoma / pathology

Substances

  • Biomarkers
  • Culture Media
  • Recombinant Proteins