Fig. 10

Functional analysis of multimodular endometrial tissue assembly in vitro: reproductive endocrine factor secretion and hormone responsiveness. The stably assembled multimodular endometrial tissue architecture was archived by inserting individually synthesized tissue blocks into multiple empty slots on the prefabricated assembly sheet. Embedded cells within individual endometrial tissue blocks were labeled with GFP (green fluorescent protein) or mCherry (red fluorescent protein) to track their incorporation into the injured sites when they were transplanted in vivo(A). For analysis of whether the multimodular endometrial tissue assembly properly produces and secretes its own reproductive endocrine factors (PGE₂ and prolactin), a cell-encapsulating tissue assembly or cell-free tissue architecture 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 evaluated by ELISAs (B). Cell-embedded tissue assembly or cell-free tissue architecture was incubated in specific culture media for 7 days and then switched from serum-supplemented medium to serum-free media. After an additional 48 h in culture, the media were harvested, and the secretion patterns of various growth factors or cytokines were analyzed by performing antibody pair-based cytokine arrays. The membrane was printed with antibodies against forty different growth factors, their receptors, and cytokines (C). The Gene Expression Omnibus (GEO) database, a public data repository for high-throughput gene expression, was also analyzed to further evaluate the correlations between the eleven secreted prominent factors (basic FGF, EGFR, HGF, IGFBP-1, IGFBP-4, IGFBP-6, and VEGF-A) and various menstruation-associated endometrial changes (D). Significant differences are presented. *p < 0.05, **p < 0.005, and ***p < 0.001 (two-sample t-test)