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Isolation, Culture, and Identification of Amniotic Fluid-Derived Mesenchymal Stem Cells

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Abstract

Amniotic fluid-derived mesenchymal stem cells (AF-MSC) are newly described, excellent seed cells that have good differentiation capability and are convenient to obtain. However, it is important to develop a method to isolate and culture AF-MSC efficiently. Amniotic fluid samples were obtained from rabbits and the adherence method was used for AF-MSC culture. Flow cytometry, western blot, and immunofluorescence studies were used to analyze the phenotypic characteristics of the cultured AF-MSC. Amniotic fluid-derived mesenchymal stem cells were successfully isolated and cultured from amniotic fluid. Flow cytometric analysis demonstrated that these cells expressed CD29 and CD44, while they did not express CD34. The expression of transcription factor Oct-4 was confirmed by western blot and immunofluorescence analysis. Using the adherence method, we developed a successful, reproducible protocol for the isolation of AF-MSC from amniotic fluid. The results of our phenotypic analysis revealed that the AF-MSC isolated in the present study were multipotent cells.

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Acknowledgments

This work was supported by Nature Science Foundation of China Grant 81170124/H0203.

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Authors and Affiliations

  1. Department of Cardiovascular Diseases, School of Medicine, Xinhua Hospital, Shanghai Jiaotong University, 1665 Kongjiang Road, Shanghai, 200092, China

    Xuetao Fei, Shan Jiang, Song Zhang & Yigang Li

  2. Department of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai, 200032, China

    Junbo Ge

  3. Department of Cardiovascular Diseases, Eastern District of Renji Hospital, Shanghai Jiaotong University, 1630 Dongfang Road, Shanghai, 200127, China

    Ben He

  4. Department of Cardiology, Washington Hospital Center, 110 Irving Street, Washington, DC, 20010, USA

    Steven Goldstein & George Ruiz

  5. Taishan Medical University, 619 Changcheng Road, Taian, 271016, Shangdong, China

    Shan Jiang

  6. Department of Cardiovascular Diseases, Jungong Hospital, 15 Yufeng Road, Qinhuangdao, 066000, Hebei, China

    Xuetao Fei

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  1. Xuetao Fei
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  2. Shan Jiang
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  3. Song Zhang
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  4. Yigang Li
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  5. Junbo Ge
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  6. Ben He
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  7. Steven Goldstein
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  8. George Ruiz
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Correspondence to Song Zhang or Yigang Li.

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Shan Jiang: Co-first author.

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Fei, X., Jiang, S., Zhang, S. et al. Isolation, Culture, and Identification of Amniotic Fluid-Derived Mesenchymal Stem Cells. Cell Biochem Biophys 67, 689–694 (2013). https://doi.org/10.1007/s12013-013-9558-z

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  • DOI: https://doi.org/10.1007/s12013-013-9558-z

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