Enzymatic digestion versus collagenase I digestion combined with mechanical dissociation for isolation of endometrial stem cells

doi:10.3969/j.issn.2095-4344.2013.06.023

Abstract

Abstract:

BACKGROUND: There have been no standard methods used to isolate and culture of endometrial stem cells and the existing methods show poor repeatability.
OBJECTIVE: To search a method that can be used to conveniently, high-efficiently and scientifically isolate endometrial stem cells.
METHODS: Endometrial stem cells were isolated by collagenase Ⅰ digestion combined with mechanical dissociation and collagenase Ⅰ digestion. The number and growth cycle of living cells were compared between these two methods. CD90 and CD117 expression levels were detected by immunocytochemical staining and RT-PCR method.
RESULTS AND CONCLUSION: After 15 days of primary culture, endometrial stem cells exhibited a shuttle-shaped appearance, adhered to the wall of culture dish, showed the morphology of fibroblasts and arranged in the absence of polarity. The expression of stem cell markers CD90 and CD117 was detected by immunocytochemical staining and RT-PCR method. This suggests that the cultured cells exhibit the characteristics of stem cells. Collagenase Ⅰ digestion combined with mechanical dissociation yielded higher number of living cells than enzymatic digestion (P < 0.01), but the growth cycle was the same after cultured by these two methods. These results indicate that collagenase Ⅰ digestion combined with mechanical dissociation is a convenient, highly efficient and scientific method used for in vitro isolation of endometrial stem cells.

Key words: stem cells, stem cell culture and differentiation, endometrial stem cells, isolation, collagenase Ⅰ, mixture, mechanical dissociation, enzymatic digestion, CD90, CD117, provincial grants-supported paper, stem cells photographs-containing paper

CLC Number:

R394.2

Ma Ying, He Yuan-li. Enzymatic digestion versus collagenase I digestion combined with mechanical dissociation for isolation of endometrial stem cells[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(6): 1089-1093.

Figures/Tables 8

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