Use of Limiting Dilution Method for Isolation of Nucleus Pulposus Mesenchymal StemProgenitor Cells and Effects of Plating Density on Biological Characteristics and Plasticity
Joint Authors
Lin, Linghan
Jia, Zhiwei
Zhao, Yachao
Wu, Yaohong
Zhao, Xiyan
Li, Yong
Guo, Ziming
Chen, Jiahai
Cheng, Shi
Wang, Deli
Ruan, Dike
Source
Issue
Vol. 2017, Issue 2017 (31 Dec. 2017), pp.1-16, 16 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2017-10-08
Country of Publication
Egypt
No. of Pages
16
Main Subjects
Abstract EN
Objectives.
To evaluate the effects of the limiting dilution method and plating density in rat nucleus pulposus mesenchymal stem/progenitor cells (NPMSCs).
Materials and Methods.
Nucleus pulposus tissues were isolated from 12-week-old male Sprague-Dawley rats and NPMSCs were isolated using limiting dilution method.
Cells were then classified into 3 groups according to plating density.
Cell morphologies were observed, and colony-forming units, migration abilities, proliferative capacities, cell cycle percentages, multilineage differentiation capacities, stem cell biomarker expression levels, and immunophenotyping were also examined in each group.
Results.
Low density group (LD) had higher morphological homogeneity, stronger colony-forming ability, higher cell proliferation capacity, and enhanced cell migration ability relative to the other two groups (p<0.05).
Moreover, LD had more cells entering S phase, with fewer cells arrested in G0/G1 phase (p<0.05).
While all three density groups showed a multilineage differentiation potential, LD showed a higher degree of observed and semiquantified lineage specific staining (p<0.05).
Furthermore, LD displayed higher expression levels of stem cell biomarkers (Nanog, Oct4, and Sox2) and showed higher percentages of CD29+, CD44+, and CD90+ cells (p<0.05) following flow cytometry analysis.
Conclusions.
Limiting dilution method is suggested when isolating NPMSCs as a means of improving cell activity and plasticity.
American Psychological Association (APA)
Lin, Linghan& Jia, Zhiwei& Zhao, Yachao& Wu, Yaohong& Zhao, Xiyan& Li, Yong…[et al.]. 2017. Use of Limiting Dilution Method for Isolation of Nucleus Pulposus Mesenchymal StemProgenitor Cells and Effects of Plating Density on Biological Characteristics and Plasticity. BioMed Research International،Vol. 2017, no. 2017, pp.1-16.
https://search.emarefa.net/detail/BIM-1139721
Modern Language Association (MLA)
Lin, Linghan…[et al.]. Use of Limiting Dilution Method for Isolation of Nucleus Pulposus Mesenchymal StemProgenitor Cells and Effects of Plating Density on Biological Characteristics and Plasticity. BioMed Research International No. 2017 (2017), pp.1-16.
https://search.emarefa.net/detail/BIM-1139721
American Medical Association (AMA)
Lin, Linghan& Jia, Zhiwei& Zhao, Yachao& Wu, Yaohong& Zhao, Xiyan& Li, Yong…[et al.]. Use of Limiting Dilution Method for Isolation of Nucleus Pulposus Mesenchymal StemProgenitor Cells and Effects of Plating Density on Biological Characteristics and Plasticity. BioMed Research International. 2017. Vol. 2017, no. 2017, pp.1-16.
https://search.emarefa.net/detail/BIM-1139721
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1139721