In vitro propagation and cardiac differentiation of canine induced pluripotent stem cells on carbon nanotube substrates

Induced pluripotent stem cells (iPSCs) have attracted an interest for personalized cell based therapy along with various other applications. There have been few studies that effective nanomaterial based scaffolds act as alternative to the commonly used feeder dependent in vitro maintenance of iPSCs. The present study provides the fundamental information on ex vivo behavior of canine iPSC (ciPSCs) maintained on carboxylic acid (-COOH) functionalized single-walled carbon nanotubes (COOH-SWCNTs) and multi-walled carbon nanotubes (COOH-MWCNTs) substrates. Here in we evaluated the comparative colony morphology, propagation, characterization, cytocompatibility and differentiation capability of ciPSC cultured on MEF feeder taken as control, and COOH-SWCNTs and COOH-MWCNTs substrates. We observed a healthy growth of ciPSCs on both the types of carbon nanotubes (CNTs) similar to feeder. The ciPSC colonies grown on both CNTs were positive for alkaline phosphatase staining and expressed pluripotent markers with notable significance. Further, the ciPSC colonies grew on these CNTs retained the in vitro differentiation ability into three germ layers as well as cardiac cell. Cytotoxicity analysis revealed that (-COOH) functionalized CNTs provided a culture condition of low cytotoxicity. The results of the present study indicated that (-COOH) functionalized CNTs could be used as xeno-free substrate to support the maintenance of iPSCs.