Generation of induced pluripotent mouse stem cells in an indirect co-culture system

Abstract

ABSTRACT. Typically, production of induced pluripotent stem cells requires direct contact with feeder cells. However, once the stem cells have reached the appropriate maturation point, it is difficult to separate them from feeder cells, which must be irradiated with γ-rays or treated with the antibiotic mitomycin-C. We used a microporous poly-membrane-based indirect contact co-culture system with mouse embryonic fibroblasts to induce mouse pluripotent stem cells without radiation or antibiotics. We found that induced pluripotent stem cells induced by this co-culture method had a reprogramming efficiency and time similar to those induced using traditional methods. Furthermore, strongly expressed pluripotent markers showed a normal karyotype and formation and contained all three germ layers in a teratoma.

Non-Genetic Direct Reprogramming and Biomimetic Platforms in a Preliminary Study for Adipose-Derived Stem Cells into Corneal Endothelia-Like Cells

“…The “biomimetic’’ environments can direct the changes of stem cells. Biomimetic platforms in vitro include regulatory molecules and signals from culture condition and other cells (e.g., co-culture) [20] , extracellular matrix (ECM) environment (e.g., decellularized ECM scaffold) [21] , [22] , and physical factors (e.g., bioreactor dynamic condition) [23] , [24] , which can be established to act in concert, with synergistic and competing effects on the reprogramming and differentiation of stem cells [25] . We previously found that 1/4 suspension of iPSCs labeled with 10 nmol/L quantum dots and 60% confluence of rabbit corneal endothelial cells (CECs) showed optimal effects on mixture co-culture each other and cell labeling.…”