Journal of Integrative Agriculture ›› 2014, Vol. 13 ›› Issue (4): 837-848.DOI: 10.1016/S2095-3119(13)60497-9

• 论文 • 上一篇    下一篇

Characterization and Differentiation into Adipocytes and Myocytes of Porcine Bone Marrow Mesenchymal Stem Cells

 DU Min-qing, HUANG Yue-qin, LU Nai-Sheng, SHU Gang, ZHU Xiao-tong, WANG Li-na, GAO Ping   

  1. College of Animal Sciences, South China Agricultural University, Guangzhou 510642, P.R.China
  • 收稿日期:2012-11-13 出版日期:2014-04-01 发布日期:2014-04-16
  • 通讯作者: WANG Song-bo, Tel/Fax: +86-20-85284901, E-mail: songbowang@scau.edu.cn; JIANG Qing-yan,Tel: +86-20-85284930, E-mail: qyjiang@scau.edu.cn
  • 作者简介:DU Min-qing, E-mail: luoluoweiwei@126.com
  • 基金资助:

    This work was supported by the National Natural Science Foundation of China (30901058 and 30972157), the National Basic Research Program of China (2009CB941601), the Natural Science Foundation of Guangdong Province of China (S2012010010176 and S2012020011048), the Special Fund for Agro-Scientific Research in the Public Interest, China (201003011), and the Specialized Research Fund for the Doctoral Program of Higher Education of China (20094404120012).

Characterization and Differentiation into Adipocytes and Myocytes of Porcine Bone Marrow Mesenchymal Stem Cells

 DU Min-qing, HUANG Yue-qin, LU Nai-Sheng, SHU Gang, ZHU Xiao-tong, WANG Li-na, GAO Ping   

  1. College of Animal Sciences, South China Agricultural University, Guangzhou 510642, P.R.China
  • Received:2012-11-13 Online:2014-04-01 Published:2014-04-16
  • Contact: WANG Song-bo, Tel/Fax: +86-20-85284901, E-mail: songbowang@scau.edu.cn; JIANG Qing-yan,Tel: +86-20-85284930, E-mail: qyjiang@scau.edu.cn
  • About author:DU Min-qing, E-mail: luoluoweiwei@126.com
  • Supported by:

    This work was supported by the National Natural Science Foundation of China (30901058 and 30972157), the National Basic Research Program of China (2009CB941601), the Natural Science Foundation of Guangdong Province of China (S2012010010176 and S2012020011048), the Special Fund for Agro-Scientific Research in the Public Interest, China (201003011), and the Specialized Research Fund for the Doctoral Program of Higher Education of China (20094404120012).

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摘要： Bone marrow mesenchymal stem cells (BMSCs) could differentiate into various cell types including adipocytes and myocytes, which had important scientific significance not only in the field of tissue regeneration, but also in the field of agricultural science. In an attempt to exhibit the characterization and differentiation into adipocytes and myocytes of porcine BMSCs, we isolated and purified porcine BMSCs by red blood cell lysis method and percoll gradient centrifugation. The purified cells presented a stretched fibroblast-like phenotype when adhered to the culture plate. The results of flow cytometry analysis and immunofluorescence staining demonstrated that the isolated cells were positive for mesenchymal surface markers CD29, CD44 and negative for hematopoietic markers CD45 and the adhesion molecules CD31. Cells were induced to differentiate into adipocytes with adipogenic medium containing insulin, dexamethasone, oleate and octanoate. Oil Red O staining demonstrated that the porcine BMSCs successfully differentiated to adipocytes. Moreover, the findings of real-time PCR and Western blotting indicated that the induced cells expressed adipogenic marker genes (PPAR-γ, C/EBP-α, perilipin, aP2) mRNA or proteins (PPAR-γ, perilipin, aP2). On the other hand, porcine BMSCs were induced into myoctyes with myogenic medium supplemented with 5-azacytidine, basic fibroblast growth factor, chick embryo extract and horse serum. Morphological observation by hochest 33342 staining showed that the induced cells presented as multi-nucleus muscular tube structure. And myogenic marker genes (Myf5, desmin) mRNA or proteins (Myf5, MyoD, myogenin, desmin) were found in the induced cells. In addition, the results of immunofluorescence staining revealed that myogenic marker (Myf5, MyoD, myogenin, desmin, S-MyHC) proteins was positive in the induced cells. Above all, these results suggested that the isolated porcine BMSCs were not only consistent with the characterization of mesenchymal stem cells, but also exhibited the multipotential capacity to form adipocytes and myocytes, which provided the basis to investigate the regulation mechanism involved in the selective differentiation of porcine BMSCs.

关键词: porcine , bone marrow mesenchymal stem cell , adipocyte , myocyte

Abstract: Bone marrow mesenchymal stem cells (BMSCs) could differentiate into various cell types including adipocytes and myocytes, which had important scientific significance not only in the field of tissue regeneration, but also in the field of agricultural science. In an attempt to exhibit the characterization and differentiation into adipocytes and myocytes of porcine BMSCs, we isolated and purified porcine BMSCs by red blood cell lysis method and percoll gradient centrifugation. The purified cells presented a stretched fibroblast-like phenotype when adhered to the culture plate. The results of flow cytometry analysis and immunofluorescence staining demonstrated that the isolated cells were positive for mesenchymal surface markers CD29, CD44 and negative for hematopoietic markers CD45 and the adhesion molecules CD31. Cells were induced to differentiate into adipocytes with adipogenic medium containing insulin, dexamethasone, oleate and octanoate. Oil Red O staining demonstrated that the porcine BMSCs successfully differentiated to adipocytes. Moreover, the findings of real-time PCR and Western blotting indicated that the induced cells expressed adipogenic marker genes (PPAR-γ, C/EBP-α, perilipin, aP2) mRNA or proteins (PPAR-γ, perilipin, aP2). On the other hand, porcine BMSCs were induced into myoctyes with myogenic medium supplemented with 5-azacytidine, basic fibroblast growth factor, chick embryo extract and horse serum. Morphological observation by hochest 33342 staining showed that the induced cells presented as multi-nucleus muscular tube structure. And myogenic marker genes (Myf5, desmin) mRNA or proteins (Myf5, MyoD, myogenin, desmin) were found in the induced cells. In addition, the results of immunofluorescence staining revealed that myogenic marker (Myf5, MyoD, myogenin, desmin, S-MyHC) proteins was positive in the induced cells. Above all, these results suggested that the isolated porcine BMSCs were not only consistent with the characterization of mesenchymal stem cells, but also exhibited the multipotential capacity to form adipocytes and myocytes, which provided the basis to investigate the regulation mechanism involved in the selective differentiation of porcine BMSCs.

Key words: porcine , bone marrow mesenchymal stem cell , adipocyte , myocyte