Fig. 9

Functional evaluation of various endometrial encapsulating tissue blocks: reproductive endocrine factor secretion and steroid hormone responsiveness. For analysis of whether the embedded endometrial stem cells within a tissue block properly secrete their own reproductive endocrine factor, a cell-encapsulating block or cell-free block was incubated in specific culture media for 7 days and then switched from serum-supplemented medium into serum-free media. After an additional 48 h in culture, the media were harvested, and PGE₂ and prolactin secretion was measured by ELISAs (A). For analysis of the responsiveness of each endometrial tissue block to the steroid hormone estrogen, the cell viability and metabolic activity of embedded cells within individual tissue blocks were assessed by performing live & dead assays and CCK-8 assays, respectively. Various endometrial tissue blocks were incubated in specific culture media for 7 days with or without estrogen exposure, and then, the live & dead assay agents were added to the block samples. Cell viabilities within individual tissue blocks were assessed using a fluorescence microscope. In addition, to assess their metabolic activities, various endometrial tissue blocks were incubated in specific culture media for 7 days with or without estrogen exposure and then switched from serum-supplemented medium to serum-free media. The block samples were additionally cultured for 48 h with CCK-8 solution in serum-free media (B). All experiments were performed in triplicate. Significant differences are presented. *p < 0.05, **p < 0.005, and ***p < 0.001 (two-sample t-test)