Proliferating a large amount of human amnion mesenchymal stem cells in vitro

doi:10.3969/j.issn.2095-4344.2013.49.019

Abstract

Abstract:

BACKGROUND: Amnion mesenchymal-derived stem cells are obtained from the placenta with a placenta amniotic membrane of about 600 cm2.
OBJECTIVE: To establish a kind of simple isolation and culture method of mesenchymal stem cells derived from human amnion in vitro, and to explore their biological properties.
METHODS: Mesenchymal stem cells derived from human amnion were harvested by trypsin digestion combined with direct adherence. The morphology of human amnion-derived mesenchymal stem cells was observed. The passage 5 cells were collected to draw a cell growth curve. Surface markers and cell cycle of the passage 5 cells were determined using flow cytometry. Passage 4 cells were obtained for osteogenic and adipogenic induction. After 4 months of cryopreservation, the resuscitated passage 4 cells were counted to determine cell survival rate and draw the cell growth curve.
RESULTS AND CONCLUSION: A few of cells creped at day after primary seeding. About after 15 days, 80%-90% cells fused in a spindle shape. After passage, the cells showed even morphology and arranged spirally. The latency period of the passaged cells was 48 hours and logarithmic growth phase was about 4 days. After logarithmic growth phase, the cells entered the platform period. The flow cytometry results showed negative expression of CD34, CD14, HLA-DR, CD19, CD45, but positive expression of CD73, CD105, CD90 on the surface of mesenchymal stem cells. The alizarin red and oil red O staining was positive and confirmed osteogenic and adipogenic capacity of human amnion-derived mesenchymal stem cells. Flow cytometry results showed that 28% cells were in S phase. After cryopreservation, the survival rate of resuscitated cells was up to 90%, and the resuscitated cells had the same growth characteristic with the non-cryopreserved cells. These results confirm a simple method to proliferate a great amount of human amnion-derived mesenchymal stem cells.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: stem cells, mesenchymal stem cells, osteoblasts, amnion

CLC Number:

R394.2

Bi Wei-wei, He Li-rong, Li Chao, Nie De-zhi. Proliferating a large amount of human amnion mesenchymal stem cells in vitro[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(49): 8583-8589.

Figures/Tables 8

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