Long-term culture of human embryonic stem cells in feeder-free conditions.
We have demonstrated previously that human embryonic stem (hES) cells possess a characteristic morphologic, antigenic, and molecular profile that can be used to assess the state of ES cells (Carpenter et al., [2004] Dev Dyn 229:243-258). In this manuscript, we have examined the long-term stability of three hES cell lines in feeder-free culture. We demonstrate that the expression of antigens and transcription factors, telomerase activity, telomere length, and karyotype appear stable for all three hES cell lines after continuous culture for over 1 yr. All three lines retained pluripotent differentiation in vitro and in vivo. Although hES cell lines were remarkably stable over the period of analysis, a detailed quantitative analysis of antigen expression by flow cytometry and gene expression by microarray suggested that cell lines show subtle differences in the expression of small subsets of genes upon long-term culture.
Rosler
Elen S
ES
Geron Corporation, Menlo Park, California, USA.
Fisk
Gregory J
GJ
Ares
Ximena
X
Irving
John
J
Miura
Takumi
T
Rao
Mahendra S
MS
Carpenter
Melissa K
MK
D000818:Animals; D000954:Antigens, Surface / Q000096:biosynthesis; D002454:Cell Differentiation; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D017077:Culture Media, Conditioned; D004622:Embryo, Mammalian / Q000166:cytology; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D007621:Karyotyping; D051379:Mice; D016513:Mice, SCID; D020411:Oligonucleotide Array Sequence Analysis; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D019098:Telomerase / Q000378:metabolism; D013724:Teratoma / Q000209:etiology / Q000473:pathology; D013997:Time Factors
D000954:Antigens, Surface; D017077:Culture Media, Conditioned; D019098:Telomerase