Open Access Research

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

Nilendran Prathalingam12*, Linda Ferguson13, Lesley Young4, Georg Lietz5, Rachel Oldershaw13, Lyn Healy4, Albert Craig16, Helen Lister16, Rakesh Binaykia1, Radhika Sheth1, Alison Murdoch167 and Mary Herbert127

Author Affiliations

1 NorthEast England Stem Cell Institute, Centre for Life, Times Square, Newcastle upon Tyne NE1 4EP, UK

2 Institute for Ageing and Health, Newcastle University, Centre for Life, Times Square, Newcastle upon Tyne NE1 4EP, UK

3 Institute for Cellular Medicine, Centre for Life, Times Square, Newcastle upon Tyne NE1 4EP, UK

4 UK Stem Cell Bank, National Institute for Biological Standards and Control, Blanche Lane, South Mimms Potters Bar, Hertfordshire, EN6 3QG, UK

5 School of Agriculture, Food and Rural Development, University of Newcastle, Kings Road, Newcastle upon Tyne NE1 7RU, UK

6 Institute for Genetic Medicine, Newcastle University, Central Parkway, Times Square, Newcastle upon Tyne, NE1 4EP, UK

7 Newcastle Fertility Centre, Centre for Life, Times Square, Newcastle upon Tyne NE1 4EP, UK

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Stem Cell Research & Therapy 2012, 3:12  doi:10.1186/scrt103

Published: 28 March 2012

Abstract

Introduction

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.