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Updated: Aug 19, 2026
Isolation of Human Mesenchymal Stem Cells and their Cultivation on the Porous Bone Matrix
Published on: February 9, 2015
[Isolation, cultivation and biological identification of human fetal marrow mesenchymal stem cells]
Nian-hai He1, Wen-li Zhao, Yu-ming Wang
1Institute of Infectious Diseases of PLA, Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
Objective:
Noting the morphological and cytobiology characteristics and phenotypes of MMSCs, to establish an isolation and culture method for fetal MMSCs in order to provide a source of marrow mesenchymal stem cells (MMSCs).
Methods:
Fetal MMSCs were isolated and cultured with in vitro cell culture technique; the characteristics of the proliferating and growing fetal MMSCs were studied with MTT and image analysis; the phenotypes of MMSCs were identified by flow cytometry and immunohistochemistry.
Results:
Bone marrow of 12 fetuses was isolated within 0.5-2 h, and about 300+/-80 adherent cells were obtained at 24 h. Colonies with more than 5 cells were 15+/-6, growth detention period of culture cell was at 1-3 d after planting, log phase growth period was at day 4, and the amount of disintegration phase cells was reduced significantly. Original culture and serial subcultivations showed that cells divided prosperiously; unequal divisions special for stem cells were observed, and the amount of MMSCs harvested from each fetus was as much as 10(11)-10(12) cells after 10 serial subcultivations. The phenotype of MMSCs was CD166 positive and CD34 negative. Serial subcultivated MMSCs expressed a microamount of AFP and did not express albumine or CK18.
Conclusion:
Fetal MMSCs are easily isolated and proliferate prosperouly. Serial subcultivated MMSCs did not differentiate into hepatocyte-like cells under common culture condition and are feasibile as seed cells for tissue engineering reconstruction.
