Three-dimensional modeling of the human fallopian tube fimbriae

Abstract

Objective Ovarian cancer is the most lethal gynecological malignancy that affects women. Recent data suggests the disease may originate in the fallopian fimbriae; however, the anatomical origin of ovarian carcinogenesis remains unclear. This is largely driven by our lack of knowledge regarding the structure and function of normal fimbriae and the relative paucity of models that accurately recapitulate the in vivo fallopian tube. Therefore, a human three-dimensional (3D) culture system was developed to examine the role of the fallopian fimbriae in serous tumorigenesis. Methods Alginate matrix was utilized to support human fallopian fimbriae ex vivo. Fimbriae were cultured with factors hypothesized to contribute to carcinogenesis, namely; H2O2 (1mM) a mimetic of oxidative stress, insulin (5 µg/ml) to stimulate glycolysis, and estradiol (E2, 10nM) which peaks before ovulation. Cultures were evaluated for changes in proliferation and p53 expression, criteria utilized to identify potential precursor lesions. Further, secretory factors were assessed after treatment with E2 to identify if steroid signaling induces a pro-tumorigenic microenvironment. Results 3D fimbriae cultures maintained normal tissue architecture up to 7 days, retaining both epithelial subtypes. Treatment of cultures with H2O2 or insulin significantly induced proliferation. However, p53 stabilization was unaffected by any particular treatment, although was induced by ex vivo culturing. Moreover, E2-alone treatment significantly induced its canonical target PR and expression of IL8, a factor linked to poor outcome. Conclusions 3D alginate cultures of human fallopian fimbriae provide an important microphysicological model, which can be further utilized to investigate serous tumorigenesis originating from the fallopian tube.

Designer Self‐Assembling Peptide Hydrogels to Engineer 3D Cell Microenvironments for Cell Constructs Formation and Precise Oncology Remodeling in Ovarian Cancer

“…To capture the oncogenic drivers and tumor biology of HGSOC, fallopian tube‐based model systems may define the physiology and susceptibility of this epithelium to tumorous transformation. [ 214 ] So, much attention must be strictly paid to the human ovarian cancer cell models to explore different aspects in the tumorigenesis. In the future, as the avenues toward personalized precise medicine take a leap to explore more and more precise 3D cell models in vitro for patient‐specific therapies and wide patient‐derived cancer research, [ 144a ] immense efforts should be taken into account to explore growth conditions of particular subpopulations of tumor cells and high density coculture systems with immune cells or other cell subtypes.…”

Ovulatory follicular fluid induces sperm phagocytosis by neutrophils, but oviductal fluid around oestrus suppresses its inflammatory effect in the buffalo oviduct in vitro

“…In the same way, a recent study by our group showed that 5 ng/ml of E2 could induce the mRNA expression of IL‐8 in BOEC and stimulate tumor necrosis factor and IL‐1B (pro‐inflammatory cytokines) expression after coculture with sperm (Yousef et al, ). Moreover, Eddie et al () reported that 2.7 ng/ml of E2 upregulated the transcription of IL‐8 in human oviductal epithelial cells. Collectively, oestradiol levels in the preovulatory FF are likely to induce an inflammatory response in the bovine oviduct.…”

WOMEN IN REPRODUCTIVE SCIENCE: Lessons from bioengineering the ovarian follicle: a personal perspective

“…Follicle maturation is autonomous Theca cells differentation is autonomous to the follicle Ovulation is autonomous to the follicle Physical rigidity alone can alter the hormone synthesis ratio (increasing rigidity favors androgen over estrogen) The ovulation rupture zone is a unique site Follicles reconstruct a holostructure after rupture in an autonomous manner The granulosa to luteal transition does not require exogenous cells Mouse follicles can function on a 28-day cycle First live birth in mouse from in vitro-grown 3D follicles First human MII eggs from in vitro-grown human follicles First ovarian bioprosthetic First microfluidic system that integrates ovarian function with reproductive tract tissues Zinc biology 20 billion zinc atoms are concentrated during meiotic maturation in the egg Telophase includes a zinc-depending zinc switch Zinc is exocytosed at the time of fertilization (zinc spark) Zinc is the cytoplasmic maturation factor Zinc is the factor responsible for the fast block to polyspermy Egg biology Meiotic chromosomes are ten times stiffer than mitotic chromosomes Meiotic chromosomes become more stiff with age Genes in polar body 1 and polar body 2 are indicative of egg maturity Terms coined Oncofertility -field of medicine at the intersection of cancer and reproductive care Bioprosthesis -tissue construct that restores soft organ function; originally ovarian bioprosthetic but term appropriate to any biologically constructed tissue Repropedia -short definitions, images or videos of reproductive terms; can be linked to any website Reprotopia -aggregate site for educational tools in the reproductive sciences EVATAR -name of microfluidic system connecting ovary, fallopian tube, uterus, cervix and liver tissue constructs; functions for 28 days Zinc spark -term for exocytotic release of zinc atoms at the time of fertilization Policies addressed Inclusion of sex as a biological variable in biomedical research Economic gaps for women in science Need for rational policies on the use of human gametes in research Insurance and reimbursment for oncofertility care Coordinated oncofertility science and medicine (a duet that recirculates) and a Quintet-MFP (Xiao et al 2017). In these systems, we cultured mouse ovarian follicles and ovaries along with human fallopian tubes, uterus, cervix and liver organoids (Laronda et al 2013, Eddie et al 2014, Arslan et al 2015, Eddie et al 2015, Zhu et al 2016, Olalekan et al 2017. This was a fascinating project and it took 5 years to invent a device that could support tissue-level function using fluidic channels that could manage the long-term (over 30 days), accurate movement of media between tissues.…”