The Isolation and Evaluation of Endometrial Epithelial Cells using the Hydrolysis at Room Temperature Combined with Differential Centrifugation

Objective To explore the effect of room temperature enzymolysis combined with improved differential centrifugation on the isolation and culture of human endometrial glandular epithelial cells and the evaluation of cell viability. Methods Endometrium specimens were collected at late proliferative and secretory phase from 10 patients with endometriosis. Each sample was divided into two for cell isolation via room temperature enzymolysis combined with improved differential centrifugation( improved group) or 37 ℃ water bath digestion combined with mesh method( control group). The number,purity,and viability of the isolated cells were compared by immunocytochemical staining and MTT assay. Results The cell isolation and culture was successful for all specimens. We obtained( 9 ± 1. 7) × 107gland epithelial cells per 1g endometrial tissue in the improved group,as compared with( 3. 9 ± 0. 78) × 107cells in the control group. The immunohistochemistry revealed that the expression of cytokeratin was positive for both groups and the purity for the improved and control group was( 97. 8 ± 0. 002) % and( 88. 6 ± 0. 006) %,respectively( P 0. 05). The MTT assay showed significant difference in cell viability between two groups during logarithmic phase( P 0. 05). Conclusion The room temperature enzymolysis combined with improved differential centrifugation can separate high purity endometrial glandular epithelial cells with a large quantity,good viability,and long growth cycle so as to be applied in the research of reproductive physiology of endometrial disease intervention.