Effect of enzymatic and non-enzymatic isolation methods on the differentiation potential of endometrial mesenchymal stem cells

Abstract

Adult stem cells reside in various organs and tissues, including the bone marrow, heart, skin, adipose tissue, and endometrium, contributing to tissue maintenance and repair. The human endometrium undergoes cyclic proliferation, differentiation, breakdown, and shedding during a woman's reproductive life, where endometrial stem cells, such as endometrial mesenchymal stem cells (eMSCs), may be pivotal in treating gynecological disorders. This study aims to compare the effects of enzymatic and non-enzymatic isolation methods on eMSC differentiation into adipogenic, chondrogenic, osteogenic, and decidua cell lineages using immunocytochemistry and RT-PCR. Analysis of eMSCs isolated from the endometrial tissues of three healthy women using non-enzymatic and enzymatic (trypsin and collagenase type I) digestion methods for the presence of mesenchymal (CD29, CD44, CD73, CD90, CD105, ITGβ1, PDGFR, and W5C5) and hematopoietic (CD14, CD31, CD34, and CD45) stem cell markers showed no significant differences in the expression of stem cell markers that is attributable to the isolation technique. However, the trypsin enzymatic isolation technique was superior in promoting the differentiation of eMSC to osteoblast, whereas the non-enzymatic isolation method more efficiently facilitated adipocyte, chondrocyte, and decidua cell differentiation of eMSCs. These findings emphasize the importance of selecting the appropriate isolation method for eMSC studies, as the choice can significantly influence the differentiation outcomes, with potential implications for future therapeutic applications in regenerative medicine that use eMSCs as the source.

Keywords: Chondrogenic; Decidua cell differentiation; Human endometrial mesenchymal stem cells; In vitro adipogenic; Isolation methods; Osteogenic.