期刊
EMBO MOLECULAR MEDICINE
卷 5, 期 1, 页码 10-17出版社
WILEY-BLACKWELL
DOI: 10.1002/emmm.201201973
关键词
clinical grade stem cells; clinical grade human embryonic stem cell validation and banking; HLA typing; human embryonic stem cells; single cell analyses
资金
- UK Medical Research Council [G0701172, G0801061]
- MRC [G0801061, G0701172] Funding Source: UKRI
- Medical Research Council [G0801061, G0701172, G0600698B] Funding Source: researchfish
Here, we describe a pre-derivation embryo haplotyping strategy that we developed in order to maximize the efficiency and minimize the costs of establishing banks of clinical grade hESC lines in which human leukocyte antigen (HLA) haplotypes match a significant proportion of the population. Using whole genome amplification followed by medium resolution HLA typing using PCR amplification with sequence-specific primers (PCR-SSP), we have typed the parents, embryos and hESC lines from three families as well as our eight clinical grade hESC lines and shown that this technical approach is rapid, reliable and accurate. By employing this pre-derivation strategy where, based on HLA match, embryos are selected for a GMP route on day 34 of development, we would have drastically reduced our cGMP laboratory running costs.
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