Embryonic Stem Cell Laboratories, Guy's Assisted Conception Unit, Division of Women's Health, King's College School of Medicine, London, UK.
EMBO Mol Med. 2013 Jan;5(1):10-7. doi: 10.1002/emmm.201201973. Epub 2012 Nov 19.
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 3-4 of development, we would have drastically reduced our cGMP laboratory running costs.
在这里,我们描述了一种胚胎前衍生单体型分析策略,我们开发该策略的目的是为了提高建立临床级 hESC 系的效率并降低成本,这些细胞系的人类白细胞抗原(HLA)单体型与人群中的很大一部分相匹配。我们使用全基因组扩增,然后使用序列特异性引物的 PCR 扩增进行中分辨率 HLA 分型(PCR-SSP),对来自三个家庭的父母、胚胎和 hESC 系以及我们的八个临床级 hESC 系进行了分型,结果表明该技术方法快速、可靠、准确。通过采用这种预衍生策略,根据 HLA 匹配,在胚胎发育的第 3-4 天选择进行 GMP 路线,我们将大幅降低我们的 cGMP 实验室运行成本。