Fig. 7: FSH treatment significantly suppresses various tissue repair capacities of endometrial stem cells in vivo.

A schematic diagram of the overall experimental protocols as described in the ‘Materials and Methods’ section is presented (a). Mice were intravenously treated with FSH (100 IU/mouse daily for 7 consecutive days). Endometrial stem cells were then isolated from endometrial tissues using our collagenase-based primary culture method. After isolation, mouse endometrial stem cells were cultured in vitro either under continuous FSH (30 IU/ml) treatment or non-FSH treatment conditions to properly mimic the in vivo environment of FSH exposure. Subsequent inhibition of cell proliferation was assessed by MTT assays (b). FSH-mediated suppression of migration capacity in vivo was then measured using Transwell assays (c) and western blotting for MMP-2 and MMP-9 (d). FSH-mediated suppression effects on adipocyte (e) and osteoblast (f) differentiation in vivo were assessed by oil red O staining and alizarin red S staining, respectively. The relative quantification of calcium deposition and lipid droplet (LD) secretion from differencing cells was performed by measuring the absorbance of solubilized cells at 500 nm and 570 nm, respectively. Uterine endometrial tissue samples from FSH-treated or nontreated mice were collected and then fixed in 10% buffered formalin for 48 h. Paraffin sections were then stained with hematoxylin and eosin (H&E) solution. Histological evaluation showed that the functional layer of endometrial tissues was markedly reduced by consecutive FSH exposure in vivo (g). β-Actin was used as an internal control to normalize protein expression. All experiments were performed in triplicate. Data are presented as the mean ± standard deviation (SD). *p < 0.05; **p < 0.005; and ***p < 0.001 (two-sample t test).