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Isolation murine mesenchymal stem cells by positive selection

In Vitro Cellular & Developmental Biology - Animal volume 43, pages 276–282 (2007)Cite this article

Abstract

Isolation and purification of mesenchymal stem cells (MSCs) from mouse via plastic adherent cultures is arduous because of the unwanted growth of hematopoietic cells and non-MSCs. In this work, homogenous populations of CD34+ MSCs from mouse bone marrow were isolated via positive selection. For this purpose, C57Bl/6 mice were killed and bone marrow cells were aspirated before incubation with magnetic bead conjugated to anti-CD34 antibody. A sample of positively selected CD34+ cells were prepared for flow cytometry to examine the expression of CD34 antigen and others were subcultured in a 25-cm2 culture flask. To investigate the mesenchymal nature, the plastic adherent cultivated cells were induced to differentiate along osteoblastic and adipogenic lineages. Furthermore, the expression of some surface markers was investigated by flow cytometry. According to the result, purified populations of fibroblast-like CD34+ cells were achieved in the first passage (1 wk after culture initiation). The cells expressed CD34, CD44, Sca-1, and Vcam-1 antigens (markers) but not CD11b and CD45. They were capable of differentiating into osteocytes and adipocytes. This study indicated that our protocol can result in the efficient isolation of homogenous populations of MSCs from C57BL/6 mouse bone marrow. We have shown that murine bone marrow-derived CD34+ cells with plastic adherent properties and capability of differentiating into skeletal lineages in vitro are MSCs.

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Acknowledgments

We are grateful to Mr. Mohammad Fallahi-Sichani for his help in editing this article. This work was supported by Stem Cell Technology Company.

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Authors and Affiliations

  1. Stem Cells Department, Stem Cell Technology, Co, Tehran, Iran

    Samad Nadri

  2. Biochemistry Department, Faculty of Science, Payam Noor University Unit Tehran, Tehran, Iran

    Samad Nadri

  3. Hematology Department, Faculty of Medical Science, Tarbiat Modares University, P.O. Box 14115-111, Tehran, Iran

    Masoud Soleimani

Authors
  1. Samad Nadri
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  2. Masoud Soleimani
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Correspondence to Masoud Soleimani.

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Editor: J. Denry Sato

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Nadri, S., Soleimani, M. Isolation murine mesenchymal stem cells by positive selection. In Vitro Cell.Dev.Biol.-Animal 43, 276–282 (2007). https://doi.org/10.1007/s11626-007-9041-5

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  • DOI: https://doi.org/10.1007/s11626-007-9041-5

Keywords

  • Positive selection
  • Mesenchymal stem cells
  • Murine bone marrow