Open AccessKaryotypic Stability, Genotyping, Differentiation, Feeder-Free Maintenance, and Gene Expression Sampling in Three Human Embryonic Stem Cell Lines Derived Prior to August 9, 2001
Author(s) -
Sandii N. Brimble,
Xianmin Zeng,
Deborah A. Weiler,
Yongquan Luo,
Ying Li,
Ian Lyons,
William J. Freed,
Allan J. Robins,
Mahendra S. Rao,
Thomas C. Schulz
Publication year - 2004
Publication title -
stem cells and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 114
eISSN - 1557-8534
pISSN - 1547-3287
DOI - 10.1089/scd.2004.13.585
Subject(s) - biology , endoderm , embryonic stem cell , induced pluripotent stem cell , stem cell , cellular differentiation , mesoderm , karyotype , genetics , microbiology and biotechnology , ectoderm , gene , chromosome
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 feederfree 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.