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

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 r...

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Bibliographic Details
Published in:Cell biochemistry and biophysics 2013-11, Vol.67 (2), p.689-694
Main Authors: Fei, Xuetao, Jiang, Shan, Zhang, Song, Li, Yigang, Ge, Junbo, He, Ben, Goldstein, Steven, Ruiz, George
Format: Article
Language:English
Subjects:
Amniotic fluid
Amniotic Fluid - cytology
Animals
Biochemistry
Biological and Medical Physics
Biomarkers - metabolism
Biomedical and Life Sciences
Biophysics
Biotechnology
Cell Biology
Cell Culture Techniques - economics
Cell Culture Techniques - methods
Cell Separation - methods
Female
Life Sciences
Mesenchymal Stromal Cells - cytology
Mesenchymal Stromal Cells - metabolism
Octamer Transcription Factor-3 - metabolism
Original Paper
Pharmacology/Toxicology
Pregnancy
Rabbits
Reproducibility of Results
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Summary: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.
ISSN:1085-9195
1559-0283
DOI:10.1007/s12013-013-9558-z