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Comparisons of Rabbit Bone Marrow Mesenchymal Stem Cell Isolation and Culture Methods In Vitro

Bone marrow mesenchymal stem cells (BMSCs) have great potential in tissue engineering and clinical therapy, and various methods for isolation and cultivation of BMSCs have been reported. However, the best techniques are still uncertain. Therefore, we sought the most suitable among the four most comm...

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Autores principales: Zhang, Weidong, Zhang, Fangbiao, Shi, Hongcan, Tan, Rongbang, Han, Shi, Ye, Gang, Pan, Shu, Sun, Fei, Liu, Xingchen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3928292/
https://www.ncbi.nlm.nih.gov/pubmed/24558428
http://dx.doi.org/10.1371/journal.pone.0088794
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author Zhang, Weidong
Zhang, Fangbiao
Shi, Hongcan
Tan, Rongbang
Han, Shi
Ye, Gang
Pan, Shu
Sun, Fei
Liu, Xingchen
author_facet Zhang, Weidong
Zhang, Fangbiao
Shi, Hongcan
Tan, Rongbang
Han, Shi
Ye, Gang
Pan, Shu
Sun, Fei
Liu, Xingchen
author_sort Zhang, Weidong
collection PubMed
description Bone marrow mesenchymal stem cells (BMSCs) have great potential in tissue engineering and clinical therapy, and various methods for isolation and cultivation of BMSCs have been reported. However, the best techniques are still uncertain. Therefore, we sought the most suitable among the four most common methods for BMSC separation from rabbits. BMSCs were obtained from untreated whole bone marrow (BM) adherent cultures, 3 volumes of red blood cells (RBC) lysed with ammonium chloride, 6 volumes of RBC lysed with ammonium chloride, and Ficoll density gradient centrifugation. Then, isolated BMSCs were evaluated with respect to primary cell yield, number of CFU-F colonies, proliferative capacity, cell phenotype, and chondrogenic differentiation potential. Our data show that BMSCs were successfully isolated by all four methods, and each method was similar with regard to cell morphology, phenotype, and differentiation potential. However, BMSCs from untreated whole BM adherent cultures had greater primary cell yields, larger colonies, and the shortest primary culture time (P<0.05). Moreover, the 4(th) generation of cultured cells had the strongest proliferative activity, the fastest growth rate and the most numerous cells compared with other cell passage generations (P<0.05). In conclusion, untreated whole BM adherent cultures are best for rabbit BMSC isolation and the 4(th) generation of cells has the strongest proliferation capacity.
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spelling pubmed-39282922014-02-20 Comparisons of Rabbit Bone Marrow Mesenchymal Stem Cell Isolation and Culture Methods In Vitro Zhang, Weidong Zhang, Fangbiao Shi, Hongcan Tan, Rongbang Han, Shi Ye, Gang Pan, Shu Sun, Fei Liu, Xingchen PLoS One Research Article Bone marrow mesenchymal stem cells (BMSCs) have great potential in tissue engineering and clinical therapy, and various methods for isolation and cultivation of BMSCs have been reported. However, the best techniques are still uncertain. Therefore, we sought the most suitable among the four most common methods for BMSC separation from rabbits. BMSCs were obtained from untreated whole bone marrow (BM) adherent cultures, 3 volumes of red blood cells (RBC) lysed with ammonium chloride, 6 volumes of RBC lysed with ammonium chloride, and Ficoll density gradient centrifugation. Then, isolated BMSCs were evaluated with respect to primary cell yield, number of CFU-F colonies, proliferative capacity, cell phenotype, and chondrogenic differentiation potential. Our data show that BMSCs were successfully isolated by all four methods, and each method was similar with regard to cell morphology, phenotype, and differentiation potential. However, BMSCs from untreated whole BM adherent cultures had greater primary cell yields, larger colonies, and the shortest primary culture time (P<0.05). Moreover, the 4(th) generation of cultured cells had the strongest proliferative activity, the fastest growth rate and the most numerous cells compared with other cell passage generations (P<0.05). In conclusion, untreated whole BM adherent cultures are best for rabbit BMSC isolation and the 4(th) generation of cells has the strongest proliferation capacity. Public Library of Science 2014-02-18 /pmc/articles/PMC3928292/ /pubmed/24558428 http://dx.doi.org/10.1371/journal.pone.0088794 Text en © 2014 Zhang et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Zhang, Weidong
Zhang, Fangbiao
Shi, Hongcan
Tan, Rongbang
Han, Shi
Ye, Gang
Pan, Shu
Sun, Fei
Liu, Xingchen
Comparisons of Rabbit Bone Marrow Mesenchymal Stem Cell Isolation and Culture Methods In Vitro
title Comparisons of Rabbit Bone Marrow Mesenchymal Stem Cell Isolation and Culture Methods In Vitro
title_full Comparisons of Rabbit Bone Marrow Mesenchymal Stem Cell Isolation and Culture Methods In Vitro
title_fullStr Comparisons of Rabbit Bone Marrow Mesenchymal Stem Cell Isolation and Culture Methods In Vitro
title_full_unstemmed Comparisons of Rabbit Bone Marrow Mesenchymal Stem Cell Isolation and Culture Methods In Vitro
title_short Comparisons of Rabbit Bone Marrow Mesenchymal Stem Cell Isolation and Culture Methods In Vitro
title_sort comparisons of rabbit bone marrow mesenchymal stem cell isolation and culture methods in vitro
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3928292/
https://www.ncbi.nlm.nih.gov/pubmed/24558428
http://dx.doi.org/10.1371/journal.pone.0088794
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