Session: MON 561-585-Ovarian & Uterine Function II
Poster Board MON-584
Methods:We set up 14-day individual 3D cultures in our modular culture system of mouse or human cells from the fallopian tube/oviduct; uterine myometrium, and endometrium; and vaginal/cervical stroma and epithelium. Cells were either embedded in a matrix of alginate gel combined with another ECM component such as collagen or fibronectin and applied to each culture module’s surface or cultured directly on insert membranes placed within the modules. Primary follicles (95-115 μm) isolated from mice were cultured in groups of 5 in 0.5% alginate for 28 days. Follicles were treated with 10 mIU/ml FSH from day 0 to day 15 and in vitro maturation was performed on day 15. Also, we assessed the tissue-specific morphologic and hormonal responses of cultured follicles to FSH or LH and of cultured fallopian tube, uterus, and cervix/vagina tissue to estradiol (E2) or progesterone (P4).
Result: (1) Cultured mouse primary follicles responded to FSH stimulation and grew into antral stage follicles that could be in-vitro maturated and form corpora lutea. Estradiol in the follicle culture medium steadily increased in the first 15 days and dropped sharply to basal levels after in vitro maturation, followed by a peak in progesterone. Peak levels of the peptide hormones such as AMH and inhibin B appeared prior to in vitro maturation. (2) Human and mouse fallopian tissue survived for the entire 14-day culture; tissue morphology was better for the insert membrane cultures. Cilia movement was present in the tissue of both species after 14 days of culture. E2 and P4 induced OVGP1 expression in the fallopian tissues compared to controls. (3) Human endometrial cells reorganized into ball-shaped structures during 14 days in culture. ER and PR expression in the endometrial and myometrial cells correlated with E2 treatment. (4) Human stromal cells of the cervix/vagina grew in the alginate gel system after 8 days, with epithelial cells growing on top of the gel. ER and PR expression in the epithelial cells correlated with estradiol stimulation.
Conclusion: Mouse and human tissues of the female reproductive tract—ovarian follicles, fallopian tube, uterus, and vagina/cervix—can grow in our modular culture system and respond appropriately to hormonal stimulation. Future studies will attempt to connect the individual culture modules to produce a complete 3D reproductive tract that is a physiologic mimic of in vivo biology.
Nothing to Disclose: JZ, YX, YY, YL, JEB, JK, TK, TKW
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