Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (10): 1837-1840.doi: 10.3969/j.issn.1673-8225.2011.10.027

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Optimization of the isolation and cultivation method of human amniotic fluid-derived mesenchymal stem cells

Guo Su-hong1, Yuan Chun-hua2, Hou Yi-jü1, Liu Hui-ling2   

  1. 1Department of Clinical Laboratory, Jilin Medical College, Jilin  132013, Jilin Province, China
    2Department of Gynaecology and Obstetrics, Affiliated Hospital of Jilin Medical College, Jilin  132013, Jilin Province, China
  • Received:2010-10-11 Revised:2010-12-04 Online:2011-03-05 Published:2011-03-05
  • About author:Guo Su-hong★, Master, Associate professor, Department of Clinical Laboratory, Jilin Medical College, Jilin 132013, Jilin Province, China suhong725@sina.com
  • Supported by:

    the Grant for Science Technology Research Program of the Department of Education of Jilin Province during the 11th Five-Year Plan Period, No. 2007-435*

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Abstract:

BACKGROUND: There are different methods of in vitro isolating culture for human amniotic fluid-derived mesenchymal stem cells (AF-MSCs). To gain most AF-MSCs is still difficult.
OBJECTIVE: We had isolated 80 cases AF, and compared different in vitro culture methods for optimization and bolted a better method to establish foundation of AF-MSCs in clinical widespread application.
METHODS: Amniotic fluid was collected from full-term deliveries and artificial labors under aseptic condition. The amniotic fluid cells were isolated by centrifugation. The influences of different pregnancy ages, culture media, incubation densities and the time points of the first medium change on the growth of AF-MSCs were analyzed. The cell metabolism was determined and biological characteristics of AF-MSCs were observed by cytochemical staining.
RESULTS AND CONCLUSION: When the other conditions were the same, AmnioMAXⅡcomplete medium was the best medium. 5×104/cm2 was the best planting density; the best time for the first medium change was on the sixth day. The achievement ratio of amniotic fluid from fetation of intermediate stage (14-20 weeks) was higher than that of later fetation (21-36 weeks) and full-term deliveries (37-42 weeks) when the other conditions were the same. AF-MSCs highly expressed periodic acid-schiff, acid phosphatase, α-naphthol acetate esterase (α-NAE) and weakly expressed neutrophil alkaline phosphatase, but no peroxidase and Sudan black was found. Results suggest that we have found the optimized method of culturing AF-MSCs.

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