| Self-renewal of human embryonic stem cells is supported by a shortened G1 cell cycle phase. | |
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MedLine Citation:
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PMID: 16972248 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Competency for self-renewal of human embryonic stem (ES) cells is linked to pluripotency. However, there is a critical paucity of fundamental parameters of human ES cell division. In this study we show that human ES cells (H1 and H9; NIH-designated WA01 and WA09) rapidly proliferate due to a very short overall cell cycle (15-16 h) compared to somatic cells (e.g., normal diploid IMR90 fibroblasts and NT-2 teratocarcinoma cells). The human ES cell cycle maintains the four canonical cell cycle stages G1, S, G2, and M, but the duration of G1 is dramatically shortened. Bromodeoxyuridine (BrdU) incorporation and FACS analysis demonstrated that 65% of asynchronously growing human ES cells are in S phase. Immunofluorescence microscopy studies detecting BrdU labeled mitotic chromosomes, Ki67 domains, and p220(NPAT) containing Cajal bodies revealed that the durations of the S ( approximately 8 h), G2 ( approximately 4 h), and M phases ( approximately 1 h) are similar in ES and somatic cells. We determined that human ES cells remain viable after synchronization with either nocodazole or the anti-tumor drug Paclitaxel (taxol) and have an abbreviated G1 phase of only 2.5-3 h that is significantly shorter than in somatic cells. Molecular analyses using quantitative RT-PCR demonstrate that human ES cells and somatic cells express similar cell cycle markers. However, among cyclins and cyclin-dependent kinases (CDKs), we observed high mRNA levels for the G1-related CDK4 and cyclin D2 genes. We conclude that human ES cells exhibit unique G1 cell cycle kinetics and use CDK4/cyclin D2 related mechanisms to attain competency for DNA replication. |
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Authors:
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Klaus A Becker; Prachi N Ghule; Jaclyn A Therrien; Jane B Lian; Janet L Stein; Andre J van Wijnen; Gary S Stein |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural |
Journal Detail:
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Title: Journal of cellular physiology Volume: 209 ISSN: 0021-9541 ISO Abbreviation: J. Cell. Physiol. Publication Date: 2006 Dec |
Date Detail:
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Created Date: 2006-10-02 Completed Date: 2007-02-07 Revised Date: 2011-03-07 |
Medline Journal Info:
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Nlm Unique ID: 0050222 Medline TA: J Cell Physiol Country: United States |
Other Details:
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Languages: eng Pagination: 883-93 Citation Subset: IM |
Copyright Information:
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(c) 2006 Wiley-Liss, Inc. |
Affiliation:
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Department of Cell Biology and Cancer Center, University of Massachusetts Medical School, 55 Lake Avenue North, Worcester, MA 01655, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Biological Markers / metabolism Cell Differentiation Cell Line Cell Proliferation* Cyclin D2 Cyclin-Dependent Kinase 4 / genetics, metabolism Cyclins / genetics, metabolism Embryonic Stem Cells / cytology, physiology* G1 Phase / physiology* Humans Karyotyping RNA, Messenger / metabolism Time Factors |
| Grant Support | |
ID/Acronym/Agency:
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GM 032010/GM/NIGMS NIH HHS; R01 GM032010-23/GM/NIGMS NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Biological Markers; 0/CCND2 protein, human; 0/Cyclin D2; 0/Cyclins; 0/RNA, Messenger; EC 2.7.11.22/Cyclin-Dependent Kinase 4 |
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