Derivation and propagation of human embryonic stem cell lines from frozen embryos in an animal product-free environment

Emma Stephenson, Laureen Jacquet, Cristian Miere, Victoria Wood, Neli Kadeva, Glenda Cornwell, Stefano Codognotto, Yaser Dajani, Peter Braude, Dusko Ilic

Research output: Contribution to journalArticlepeer-review

61 Citations (Scopus)

Abstract

The protocols described here are comprehensive instructions for deriving human embryonic stem (hES) cell lines in xeno-free conditions from cryopreserved embryos. Details are included for propagation, cryopreservation and characterization. Initial derivation is on feeder cells and is followed by adaptation to a feeder-free environment; competent technicians can perform these simplified methods easily. From derivation to cryopreservation of fully characterized initial stocks takes 3-4 months. These protocols served as the basis for standard operating procedures (SOPs), with both operational and technical components, that we set to meet good manufacturing practice (GMP) and UK regulatory body requirements for derivation of clinical-grade cells. As such, these SOPs are currently used in our current GMP compliant facility to derive hES cell lines ab initio, in an animal product-free environment; these lines are suitable for research and potentially for clinical use in cell therapy. So far, we have derived eight clinical-grade lines, which will be freely available to the scientific community after submission/accession to the UK Stem Cell Bank.

Original languageEnglish
Pages (from-to)1366-1381
Number of pages16
JournalNature Protocols
Volume7
Issue number7
Early online date21 Jun 2012
DOIs
Publication statusPublished - Jul 2012

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