title: Karyotypic stability, genotyping, differentiation, feeder-free maintenance, and gene expression sampling in three human embryonic stem cell lines derived prior to August 9, 2001.
abstract: 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.
authors.affiliation: BresaGen, Inc., Athens, GA 30605, USA.
authors.forename: Sandii N
authors.initials: SN
authors.lastname: Brimble
authors.forename: Xianmin
authors.initials: X
authors.lastname: Zeng
authors.forename: Deborah A
authors.initials: DA
authors.lastname: Weiler
authors.forename: Yongquan
authors.initials: Y
authors.lastname: Luo
authors.forename: Ying
authors.initials: Y
authors.lastname: Liu
authors.forename: Ian G
authors.initials: IG
authors.lastname: Lyons
authors.forename: William J
authors.initials: WJ
authors.lastname: Freed
authors.forename: Allan J
authors.initials: AJ
authors.lastname: Robins
authors.forename: Mahendra S
authors.initials: MS
authors.lastname: Rao
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Schulz
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D015925:Cryopreservation; D003582:Cytogenetics / Q000379:methods; D018076:DNA, Complementary / Q000378:metabolism; D004622:Embryo, Mammalian / Q000166:cytology*; D005786:Gene Expression Regulation; D005838:Genotype; D006650:Histocompatibility Testing; D006801:Humans; D007621:Karyotyping; D008856:Microscopy, Fluorescence; D020411:Oligonucleotide Array Sequence Analysis; D039904:Pluripotent Stem Cells / Q000166:cytology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D015398:Signal Transduction; D013234:Stem Cells / Q000166:cytology*
chemical_list: D018076:DNA, Complementary; D000469:Alkaline Phosphatase