Karyotypic stability, genotyping, differentiation, feeder-free maintenance, and gene expression sampling in three human embryonic stem cell lines derived prior to August 9, 2001
Karyotypic stability, genotyping, differentiation, feeder-free maintenance, and gene expression sampling in three human embryonic stem cell lines derived prior to August 9, 2001
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2001 年 8 月 9 日之前提取的三种人类胚胎干细胞系的核型稳定性、基因分型、分化、无饲养维持和基因表达取样
DOI：
10.1089/scd.2004.13.585
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发表时间：
2004-12-01
影响因子：
4
通讯作者：
Schulz, TC
中科院分区：
文献类型：
--
作者：
Brimble, SN;Zeng, XM;Schulz, TC
美国联邦政府资助的研究人员可获取的人类胚胎干细胞（hESC）系数量目前有限。因此，确定其基本特性并传播这些细胞系非常重要。在本报告中，我们恢复并扩增了最早可获取的BG01、BG02和BG03人类胚胎干细胞系的冷冻储存样本。这些培养物展现出未分化细胞的多种明确特征，包括长期自我更新、多能性标志物的表达、正常核型的维持以及向中胚层、内胚层和外胚层的分化。每个细胞系都呈现出独特的基因型和人类白细胞抗原（HLA）同种型，证实它们是独立分离的。在无饲养层条件下维持的BG01、BG02和BG03表现出自我更新、正常核型的维持以及表明未分化多能干细胞的基因表达。利用大规模平行信号测序对BG02细胞的基因表达进行的一项调查产生了转录本丰度的数字读数，并表明该细胞系与其他人类胚胎干细胞系相似。因此，BG01、BG02和BG03人类胚胎干细胞是独立的、未分化的且多能的细胞系，能够在不积累核型异常的情况下得以维持。
The number of human embryonic stem cell (hESC) lines available to federally funded U. S. researchers is currently limited. Thus, determining their basic characteristics and disseminating these lines is important. In this report, we recovered and expanded the earliest available cryopreserved stocks of the BG01, BG02, and BG03 hESC lines. These cultures exhibited multiple definitive characteristics of undifferentiated cells, including long-term self-renewal, expression of markers of pluripotency, maintenance of a normal karyotype, and differentiation to mesoderm, endoderm, and ectoderm. Each cell line exhibited a unique genotype and human leukocyte antigen (HLA) isotype, confirming that they were isolated independently. BG01, BG02, and BG03 maintained in feeder-free conditions demonstrated self-renewal, maintenance of normal karyotype, and gene expression indicative of undifferentiated pluripotent stem cells. A survey of gene expression in BG02 cells using massively parallel signature sequencing generated a digital read-out of transcript abundance and showed that this line was similar to other hESC lines. BG01, BG02, and BG03 hESCs are therefore independent, undifferentiated, and pluripotent lines that can be maintained without accumulation of karyotypic abnormalities.