Surface-engineered substrates for improved human pluripotent stem cell culture under fully defined conditions
2011
Saha, Krishanu | Mei, Ying | Reisterer, Colin M. | Pyzocha, Neena Kenton | Yang, Jing | Muffat, Julien | Davies, Martyn C. | Alexander, Morgan R. | Langer, Robert | Anderson, Dan (Dan G.) | Jaenisch, Rudolf
The current gold standard for the culture of human pluripotent stem cells requires the use of a feeder layer of cells. Here, we develop a spatially defined culture system based on UV/ozone radiation modification of typical cell culture plastics to define a favorable surface environment for human pluripotent stem cell culture. Chemical and geometrical optimization of the surfaces enables control of early cell aggregation from fully dissociated cells, as predicted from a numerical model of cell migration, and results in significant increases in cell growth of undifferentiated cells. These chemically defined xeno-free substrates generate more than three times the number of cells than feeder-containing substrates per surface area. Further, reprogramming and typical gene-targeting protocols can be readily performed on these engineered surfaces. These substrates provide an attractive cell culture platform for the production of clinically relevant factor-free reprogrammed cells from patient tissue samples and facilitate the definition of standardized scale-up friendly methods for disease modeling and cell therapeutic applications.
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