Linked Life Data

21132465

Source:http://linkedlifedata.com/resource/pubmed/id/21132465

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2011-2-21
pubmed:abstractText
The objective of this study is to establish the aging model of murine hematopoietic stem cell (HSC) ex vitro and investigate its relative biological mechanism, aimed to build the foundation for searching the methods to delaying HSC aging. Sca-1(+)HSC were isolated and purified from murine bone marrow mono-nucleated cell by magnetic-activated cell sorting. The purity of separated cells was analyzed by flow cytometry (FCM) and the expression of Sca-1 was detected by immunofluorescence. Sca-1(+)HSC induced aging by tert-butylhydroperoxide (t-BHP, final concentration of 100 ?mol/L) for 6 h to establish the murine HSC aging model in vitro. Biological characteristics of aging HSC were evaluated by mixed hematopoietic progenitor cell culture in vitro, cell cycle assay and senescence-associated ?-galactosidase (SA-?-gal) cytochemical staining. Telomere length and telomerase activity were detected by southern blotting and telomere repeat amplification protocol-polymerase chain reaction (TRAP-PCR) augmentation. The expressions of p16(INK4a), P19(Arf), P53, P21(Cip1/Waf1) mRNA were detected by reverse transcription (RT)-PCR. The purity of separated Sca-1(+) HSC was 87.2% and the survival of Sca-1(+) HSC was 96~99%. After 6 h cocultured with 100 ?mol/L t-BHP, the ability of aging Sca-1(+) HSC to form mixed hematopoietic progenitor colony, self-renewal and multi-differentiation were decreased significantly. The number of aging Sca-1(+) HSC entered G1 phase of the cell cycle and the percentage of positive cells expressed SA-?-gal increased significantly. The telomere length shortened and the telomerase activity decreased. The expression of p16(INK4a), p19(Arf), p53, P21(Cip1/Waf1) mRNA increased. The t-BHP can induce Sca-1(+) HSC senescence in vitro. The signal transduction pathways of p16(INK4a)-retinoblastoma and P19(Arf)-Mdm2-P53-P21(Cip1/Waf1) may play key roles in the Sca-1(+) HSC senescence induced by t-BHP.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
1543-706X
pubmed:author
pubmed:issnType
Electronic
pubmed:volume
47
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
149-56
pubmed:meshHeading
pubmed:year
2011
pubmed:articleTitle
Establishment of an aging model of Sca-1+ hematopoietic stem cell and studies on its relative biological mechanisms.
pubmed:affiliation
Laboratory of Stem Cell and Tissue Engineering, Department of Histology and Embryology, Chongqing Medical University, Chongqin, China.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't