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Cell Biochemistry and Biophysics

, Volume 67, Issue 2, pp 689–694 | Cite as

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

  • Xuetao Fei
  • Shan Jiang
  • Song ZhangEmail author
  • Yigang LiEmail author
  • Junbo Ge
  • Ben He
  • Steven Goldstein
  • George Ruiz
Original Paper

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.

Keywords

Amniotic fluid-derived mesenchymal stem cells Isolation Culture Oct-4 Western blot 

Notes

Acknowledgments

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

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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Xuetao Fei
    • 1
    • 6
  • Shan Jiang
    • 1
    • 5
  • Song Zhang
    • 1
    Email author
  • Yigang Li
    • 1
    Email author
  • Junbo Ge
    • 2
  • Ben He
    • 3
  • Steven Goldstein
    • 4
  • George Ruiz
    • 4
  1. 1.Department of Cardiovascular Diseases, School of Medicine, Xinhua HospitalShanghai Jiaotong UniversityShanghaiChina
  2. 2.Department of Cardiovascular Diseases, Zhongshan HospitalFudan UniversityShanghaiChina
  3. 3.Department of Cardiovascular Diseases, Eastern District of Renji HospitalShanghai Jiaotong UniversityShanghaiChina
  4. 4.Department of CardiologyWashington Hospital CenterWashingtonUSA
  5. 5.Taishan Medical UniversityTaianChina
  6. 6.Department of Cardiovascular DiseasesJungong HospitalQinhuangdaoChina

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