Introduction: The development of reproducible methods for deriving human embryonic stem cell (hESC) lines in\r\ncompliance with good manufacturing practice (GMP) is essential for the development of hESC-based therapies.\r\nAlthough significant progress has been made toward the development of chemically defined conditions for the\r\nmaintenance and differentiation of hESCs, efficient derivation of new hESCs requires the use of fibroblast feeder\r\ncells. However, GMP-grade feeder cell lines validated for hESC derivation are not readily available.\r\nMethods: We derived a fibroblast cell line (NclFed1A) from human foreskin in compliance with GMP standards.\r\nConsent was obtained to use the cells for the production of hESCs and to generate induced pluripotent stem cells\r\n(iPSCs). We compared the line with a variety of other cell lines for its ability to support derivation and self-renewal\r\nof hESCs.\r\nResults: NclFed1A supports efficient rates (33%) of hESC colony formation after explantation of the inner cell mass\r\n(ICM) of human blastocysts. This compared favorably with two mouse embryonic fibroblast (MEF) cell lines.\r\nNclFed1A also compared favorably with commercially available foreskin fibroblasts and MEFs in promoting\r\nproliferation and pluripotency of a number of existing and widely used hESCs. The ability of NclFed1A to maintain\r\nself-renewal remained undiminished for up to 28 population doublings from the master cell bank.\r\nConclusions: The human fibroblast line Ncl1Fed1A, produced in compliance with GMP standards and qualified for\r\nderivation and maintenance of hESCs, is a useful resource for the advancement of progress toward hESC-based\r\ntherapies in regenerative medicine.
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