title: Production and validation of a good manufacturing practice grade human fibroblast line for supporting human embryonic stem cell derivation and culture.
abstract: BACKGROUND
  The development of reproducible methods for deriving human embryonic stem cell (hESC) lines in compliance with good manufacturing practice (GMP) is essential for the development of hESC-based therapies. Although significant progress has been made toward the development of chemically defined conditions for the maintenance and differentiation of hESCs, efficient derivation of new hESCs requires the use of fibroblast feeder cells. However, GMP-grade feeder cell lines validated for hESC derivation are not readily available.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: NorthEast England Stem Cell Institute, Centre for Life, Times Square, Newcastle upon Tyne NE1 4EP, UK. n.prathalingam@ncl.ac.uk
authors.forename: Nilendran
authors.initials: N
authors.lastname: Prathalingam
authors.forename: Linda
authors.initials: L
authors.lastname: Ferguson
authors.forename: Lesley
authors.initials: L
authors.lastname: Young
authors.forename: Georg
authors.initials: G
authors.lastname: Lietz
authors.forename: Rachel
authors.initials: R
authors.lastname: Oldershaw
authors.forename: Lyn
authors.initials: L
authors.lastname: Healy
authors.forename: Albert
authors.initials: A
authors.lastname: Craig
authors.forename: Helen
authors.initials: H
authors.lastname: Lister
authors.forename: Rakesh
authors.initials: R
authors.lastname: Binaykia
authors.forename: Radhika
authors.initials: R
authors.lastname: Sheth
authors.forename: Alison
authors.initials: A
authors.lastname: Murdoch
authors.forename: Mary
authors.initials: M
authors.lastname: Herbert
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002460:Cell Line / Q000166:cytology*; D064987:Cell- and Tissue-Based Therapy / Q000379:methods; D053595:Embryonic Stem Cells / Q000166:cytology*; D061252:Feeder Cells / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology; D051379:Mice; D044968:Regenerative Medicine