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Establishment of a method for isolation and cultivation of mesenchymal stem cells from human bone marrow

Xiao Liu

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Abstract

Objective: To establish a method for isolation and cultivation of mesenchymal stem cells (MSCs) from human bone marrow. Methods: Human bone marrow mononuclear cells were separated by gradient centrifugation on Percoll (density 1.073 g/ml) or Ficoll Hypaque (1.077 g/ml). The cells were incubated in DMEM (low glucose) with 10% or 20% newborn bovine serum from selected lots. The purity of MSCs was analyzed by flow cytometry. Results: The purity of MSCs collected by Percoll and cultured in medium containing 10% NBS was around 95% as assessed by flow cytometry.The purity of MSCs was much lower, nevertheless, when the cells separated by Ficoll centrifugation and / or cultured in medium with 20% NBS. Conclusions: A stable and practical method was established for separation and subsequent cultivation of human bone marrow mesenchymal stem cells. [

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Objective: To establish a method for isolation and cultivation of mesenchymal stem cells (MSCs) from human bone marrow. Methods: Human bone marrow mononuclear cells were separated by gradient centrifugation on Percoll (density 1.073 g/ml) or Ficoll Hypaque (1.077 g/ml). The cells were incubated in DMEM (low glucose) with 10% or 20% newborn bovine serum from selected lots. The purity of MSCs was analyzed by flow cytometry. Results: The purity of MSCs collected by Percoll and cultured in medium containing 10% NBS was around 95% as assessed by flow cytometry.The purity of MSCs was much lower, nevertheless, when the cells separated by Ficoll centrifugation and / or cultured in medium with 20% NBS. Conclusions: A stable and practical method was established for separation and subsequent cultivation of human bone marrow mesenchymal stem cells. [

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Available abstract

Objective: To establish a method for isolation and cultivation of mesenchymal stem cells (MSCs) from human bone marrow. Methods: Human bone marrow mononuclear cells were separated by gradient centrifugation on Percoll (density 1.073 g/ml) or Ficoll Hypaque (1.077 g/ml). The cells were incubated in DMEM (low glucose) with 10% or 20% newborn bovine serum from selected lots. The purity of MSCs was analyzed by flow cytometry. Results: The purity of MSCs collected by Percoll and cultured in medium containing 10% NBS was around 95% as assessed by flow cytometry.The purity of MSCs was much lower, nevertheless, when the cells separated by Ficoll centrifugation and / or cultured in medium with 20% NBS. Conclusions: A stable and practical method was established for separation and subsequent cultivation of human bone marrow mesenchymal stem cells. [

Key concepts: Percoll, Ficoll, Mesenchymal stem cell, Bone marrow, Flow cytometry, Centrifugation, Differential centrifugation, Human bone

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