Document Detail


Self-renewal of human embryonic stem cells is supported by a shortened G1 cell cycle phase.
MedLine Citation:
PMID:  16972248     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
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:
Type:  Journal Article; Research Support, N.I.H., Extramural    
Journal Detail:
Title:  Journal of cellular physiology     Volume:  209     ISSN:  0021-9541     ISO Abbreviation:  J. Cell. Physiol.     Publication Date:  2006 Dec 
Date Detail:
Created Date:  2006-10-02     Completed Date:  2007-02-07     Revised Date:  2014-09-11    
Medline Journal Info:
Nlm Unique ID:  0050222     Medline TA:  J Cell Physiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  883-93     Citation Subset:  IM    
Copyright Information:
(c) 2006 Wiley-Liss, Inc.
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MeSH Terms
Descriptor/Qualifier:
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:
GM 032010/GM/NIGMS NIH HHS; R01 GM032010/GM/NIGMS NIH HHS; R01 GM032010-23/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Biological Markers; 0/CCND2 protein, human; 0/Cyclin D2; 0/Cyclins; 0/RNA, Messenger; EC 2.7.11.22/Cyclin-Dependent Kinase 4

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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