Characterization and expression of senescence marker in prolonged passages of rat bone marrow-derived mesenchymal stem cells

The present study is aimed at optimizing the in vitro culture protocol for generation of rat bone marrow- (BM-) derived mesenchymal stem cells (MSCs) and characterizing the culture-mediated cellular senescence. The initial phase of generation and characterization was conducted using the adherent cel...

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Main Authors: Ridzuan, Noridzzaida, Al Abbar, Akram, Wai, Kien Yip, Maqbool, Maryam, Ramasamy, Rajesh
Format: Article
Language:English
Published: Hindawi Publishing Corporation 2016
Online Access:http://psasir.upm.edu.my/id/eprint/55258/
http://psasir.upm.edu.my/id/eprint/55258/1/Characterization%20and%20expression%20of%20senescence%20marker%20in%20prolonged%20passages%20of%20rat%20bone%20marrow.pdf
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author Ridzuan, Noridzzaida
Al Abbar, Akram
Wai, Kien Yip
Maqbool, Maryam
Ramasamy, Rajesh
author_facet Ridzuan, Noridzzaida
Al Abbar, Akram
Wai, Kien Yip
Maqbool, Maryam
Ramasamy, Rajesh
author_sort Ridzuan, Noridzzaida
building UPM Institutional Repository
collection Online Access
description The present study is aimed at optimizing the in vitro culture protocol for generation of rat bone marrow- (BM-) derived mesenchymal stem cells (MSCs) and characterizing the culture-mediated cellular senescence. The initial phase of generation and characterization was conducted using the adherent cells from Sprague Dawley (SD) rat’s BM via morphological analysis, growth kinetics, colony forming unit capacity, immunophenotyping, and mesodermal lineage differentiation. Mesenchymal stem cells were successfully generated and characterized as delineated by the expressions of CD90.1, CD44H, CD29, and CD71 and lack of CD11b/c and CD45 markers. Upon induction, rBM-MSCs differentiated into osteocytes and adipocytes and expressed osteocytes and adipocytes genes. However, a decline in cell growth was observed at passage 4 onwards and it was further deciphered through apoptosis, cell cycle, and senescence assays. Despite the enhanced cell viability at later passages (P4-5), the expression of senescence marker, β-galactosidase, was significantly increased at passage 5. Furthermore, the cell cycle analysis has confirmed the in vitro culture-mediated cellular senescence where cells were arrested at the G0/G1 phase of cell cycle. Although the currently optimized protocols had successfully yielded rBM-MSCs, the culture-mediated cellular senescence limits the growth of rBM-MSCs and its potential use in rat-based MSC research.
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spelling upm-552582018-01-22T08:35:49Z http://psasir.upm.edu.my/id/eprint/55258/ Characterization and expression of senescence marker in prolonged passages of rat bone marrow-derived mesenchymal stem cells Ridzuan, Noridzzaida Al Abbar, Akram Wai, Kien Yip Maqbool, Maryam Ramasamy, Rajesh The present study is aimed at optimizing the in vitro culture protocol for generation of rat bone marrow- (BM-) derived mesenchymal stem cells (MSCs) and characterizing the culture-mediated cellular senescence. The initial phase of generation and characterization was conducted using the adherent cells from Sprague Dawley (SD) rat’s BM via morphological analysis, growth kinetics, colony forming unit capacity, immunophenotyping, and mesodermal lineage differentiation. Mesenchymal stem cells were successfully generated and characterized as delineated by the expressions of CD90.1, CD44H, CD29, and CD71 and lack of CD11b/c and CD45 markers. Upon induction, rBM-MSCs differentiated into osteocytes and adipocytes and expressed osteocytes and adipocytes genes. However, a decline in cell growth was observed at passage 4 onwards and it was further deciphered through apoptosis, cell cycle, and senescence assays. Despite the enhanced cell viability at later passages (P4-5), the expression of senescence marker, β-galactosidase, was significantly increased at passage 5. Furthermore, the cell cycle analysis has confirmed the in vitro culture-mediated cellular senescence where cells were arrested at the G0/G1 phase of cell cycle. Although the currently optimized protocols had successfully yielded rBM-MSCs, the culture-mediated cellular senescence limits the growth of rBM-MSCs and its potential use in rat-based MSC research. Hindawi Publishing Corporation 2016 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/55258/1/Characterization%20and%20expression%20of%20senescence%20marker%20in%20prolonged%20passages%20of%20rat%20bone%20marrow.pdf Ridzuan, Noridzzaida and Al Abbar, Akram and Wai, Kien Yip and Maqbool, Maryam and Ramasamy, Rajesh (2016) Characterization and expression of senescence marker in prolonged passages of rat bone marrow-derived mesenchymal stem cells. Stem Cell International. art. no. 8487264. pp. 1-14. ISSN 1687-9678 10.1155/2016/8487264
spellingShingle Ridzuan, Noridzzaida
Al Abbar, Akram
Wai, Kien Yip
Maqbool, Maryam
Ramasamy, Rajesh
Characterization and expression of senescence marker in prolonged passages of rat bone marrow-derived mesenchymal stem cells
title Characterization and expression of senescence marker in prolonged passages of rat bone marrow-derived mesenchymal stem cells
title_full Characterization and expression of senescence marker in prolonged passages of rat bone marrow-derived mesenchymal stem cells
title_fullStr Characterization and expression of senescence marker in prolonged passages of rat bone marrow-derived mesenchymal stem cells
title_full_unstemmed Characterization and expression of senescence marker in prolonged passages of rat bone marrow-derived mesenchymal stem cells
title_short Characterization and expression of senescence marker in prolonged passages of rat bone marrow-derived mesenchymal stem cells
title_sort characterization and expression of senescence marker in prolonged passages of rat bone marrow-derived mesenchymal stem cells
url http://psasir.upm.edu.my/id/eprint/55258/
http://psasir.upm.edu.my/id/eprint/55258/
http://psasir.upm.edu.my/id/eprint/55258/1/Characterization%20and%20expression%20of%20senescence%20marker%20in%20prolonged%20passages%20of%20rat%20bone%20marrow.pdf