Defined pluripotent stem cell culture media is a valuable tool for tracking and analyzing morphological, cell signaling and gene expression changes in human embryonic stem cells. Cultivation of hESC under xeno-free culture settings provides researchers with a consistent and reproducible environment to test experimental hypotheses and move towards translational and clinical research. One of the primary concerns of the xenogeneic culture is the transfer of foreign pathogens or antigens that induce disease or immune response by the patient. These undesirable by-products may come from the use of murine-derived feeder cells, xenogeneic matrices, or from animal-derived components found in the cell culture medium or matrix used to isolate or expand the stem cells. This chapter describes standardized protocols for obtaining consistent and reproducible results for culturing PSC under xeno-free, feeder-free, or feeder-based systems.
Human embryonic stem cells can be cultured and maintained on fibroblast feeder cells of murine or human origin. Thorough protocols are provided for the growth and maintenance of human embryonic stem cells on either feeder cell type.
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