Research
Production and validation of a good manufacturing practice grade human fibroblast line for supporting human embryonic stem cell derivation and culture
- † Equal contributors
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
Stem Cell Research & Therapy 2012, 3:12 doi:10.1186/scrt103
Published: 28 March 2012Additional files
Additional file 1:
Figure S1. Images of NclFed1A showing (a) about 15% confluent, (b) about 60% confluent, and (c) 100% confluent. (d) The number of cells per square centimeter in a confluent flask was determined by counting cells with a Vi-Cell (Beckman Coulter), in eight different confluent T150 flasks. Counts were repeated 3 times for each flask.
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Additional file 2:
Table S1. PCR primers used to determine the presence of transcripts in NclFed1A that are predicted to be supportive of hESC cultures. Table S2. STR analysis with genotype copy number for allele 1 and 2. Table S3. The expression of pluripotency markers determined by FACs analysis after culture of four hESC lines for five passages on four different fibroblasts lines. Table S4. The expression of pluripotency markers determined by FACs analysis after culture of hESC lines for five passages on NclFed1A at P10, P15, P20, and P25.
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