Production and validation of a good manufacturing practice grade human fibroblast line for supporting human embryonic stem cell derivation and culture

We derived a fibroblast cell line (NclFed1A) from human foreskin in compliance with GMP standards. Consent was obtained to use the cells for the production of hESCs and to generate induced pluripotent stem cells (iPSCs). We compared the line with a variety of other cell lines for its ability to support derivation and self-renewal of hESCs.NclFed1A supports efficient rates (33%) of hESC colony formation after explantation of the inner cell mass (ICM) of human blastocysts. This compared favorably with two mouse embryonic fibroblast (MEF) cell lines. NclFed1A also compared favorably with commercially available foreskin fibroblasts and MEFs in promoting proliferation and pluripotency of a number of existing and widely used hESCs. The ability of NclFed1A to maintain self-renewal remained undiminished for up to 28 population doublings from the master cell bank.The human fibroblast line Ncl1Fed1A, produced in compliance with GMP standards and qualified for derivation and maintenance of hESCs, is a useful resource for the advancement of progress toward hESC-based therapies in regenerative medicine.Progress in the use of human embryonic stem cells (hESCs) derivatives for cellular therapies will require the production of clinical-grade lines under the control of good manufacturing practice (GMP) [1]. The ultimate goal is to increase reproducibility in the production of hESCs by developing chemically defined culture conditions by using recombinant proteins for hESC derivation and culture. A major hurdle is to dispense with the use of feeder cells, which are conventionally used to promote and support hESC self-renewal [2]. However, the ability of feeder-free culture systems to maintain genetic stability remains controversial [3]. Furthermore, reproducible techniques for deriving new GMP-grade hESC lines from human blastocysts without the use of feeder cells remain to be developed [4]. To date, only a single report describes successful hESC derivation in the absence of feeder ce