Bovine Ovarian Cortex Tissue Culture

Sutton, Courtney M; Springman, Shelby A.; Abedal-Majed, Mohamed A; Cupp, Andrea S.

doi:10.3791/61668

Cited by 5 publications

(2 citation statements)

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“…The GSCs niche is affected by a variety of signals [ 9 ]. In this niche, bone morphology protein (BMP) signaling is a key signal that regulates GSCs regeneration [ 10 ]. BMP signaling plays an important role in regulating early embryonic development, ovarian granulosa cell proliferation, oocyte maturation and reproductive hormone synthesis, depending on the ovarian paracrine/autocrine regulatory systems [ 11 ].…”

Section: Introductionmentioning

confidence: 99%

New insights into ginsenoside Rg1 regulating the niche to inhibit age-induced germline stem cells depletion through targeting ECR/BMP signaling pathway in Drosophila

Fu,

Ma,

Liu

et al. 2024

Aging

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Purpose: The age-induced imbalance in ecological niches leads to the loss of GSCs, which is the main reason for ovarian germline senescence. Ginsenoside Rg1 can delay ovarian senescence. Here, we shed light on new insights of ginsenoside Rg1 in regulating the niche to maintain GSCs self-renewal and discussing related molecular mechanisms. Methods: The differences among GSC number, reproductive capacity of naturally aging female Drosophila after ginsenoside Rg1 feeding were analyzed by immunofluorescence and behavior monitoring. The expressions of the active factors in the niche and the BMP signaling were analyzed through Western blot and RT-qPCR. The target effect was verified in the ECR mutant and combined with the molecular docking. Results: Ginsenoside Rg1 inhibited the age-induced reduction of the GSCs number and restored offspring production and development. Ginsenoside Rg1 promoted the expression of anchor proteins E-cadherin, stemness maintenance factor Nos and differentiation promoting factor Bam, thereby GSCs niche homeostasis was regulated. In addition, ginsenoside Rg1 was bound to the LBD region of the hormone receptor ECR. Ginsenoside Rg1 promotes the regeneration of GSCs by targeting the ECR to increase pSmad1/5/8 expression and thereby activating the BMP signaling pathway. In addition, ginsenoside Rg1 maintenance of niche homeostasis to promote GSCs regeneration is dependent on ECR as demonstrated in ECR mutants. Conclusions: Ginsenoside Rg1 regulated the ecological niche homeostasis of GSCs and promoted the regeneration of GSCs by targeting the ECR/BMP signaling pathway in hormone-deficient states in aging ovaries. It is of great significance for prolonging fertility potential and delaying ovarian senescence.

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Section: Introductionmentioning

confidence: 99%

New insights into ginsenoside Rg1 regulating the niche to inhibit age-induced germline stem cells depletion through targeting ECR/BMP signaling pathway in Drosophila

Fu,

Ma,

Liu

et al. 2024

Aging

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“…Furthermore, acute depletion of the follicular pool arising from over activation post grafting has been reported (Gavish et al 2014, Masciangelo et al 2019. A promising alternative to the transplantation of ovarian tissue is in vitro culture, which can be done for whole tissue (in situ) or isolated follicles (ex situ) (Gastal et al 2019, Lunardi et al 2016, O'Brien et al 2003, Paynter et al 1999, Shoorei et al 2019, Sutton et al 2021, Telfer et al 2019). In addition, tissue damage from negative effects of vitrification were found to be ameliorated during in vitro culture of warmed ovarian tissue (Meng et al 2022, Mouttham et al 2015.…”

Section: Introductionmentioning

confidence: 99%

Culture of vitrified bovine ovarian tissue on agarose gel inserts maintains follicle integrity

Alkali,

Colombo,

Luciano

et al. 2023

Reproduction

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Ovarian tissue preservation is hitherto a promising fertility insurance option for precious animals. Ovarian tissue vitrification and culture combined approach would eliminate the need of transplanting ovarian tissue to obtain mature oocytes. We aimed at optimizing vitrification and in vitro culture conditions for improved bovine ovarian tissue viability. Ovaries obtained from the slaughterhouse were punched into fragments and divided into three groups. Group 1 (fresh) was divided into two and immediately placed in two culture systems (culture inserts and agarose inserts). Group 2 was vitrified, warmed, and placed in the two culture systems while group 3 was only equilibrated then placed in the two culture systems. All cultures were maintained for six days and spent media were collected on alternate days for cytokine (interleukin 1β and interleukin 6) evaluation. Fragments were fixed for morphology assessment and immunohistochemistry. Higher percentages (P<0.05) of grade one (morphologically intact) follicles were observed in fragments on agarose compared to those on culture inserts at days two and four of culture. Conversely, we found higher (P<0.05) shifts of primordial follicles to transitional follicles in fragments on culture inserts vis-à-vis agarose inserts which was consistent with higher proportion of Ki-67 and MCM-7 and activated caspase-3 positive follicles. In conclusion, in vitro culture of bovine ovarian tissue on agarose inserts maintained follicle morphology, low follicle activation and low apoptosis compared to culture inserts.

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VEGFA165 can rescue excess steroid secretion, inflammatory markers, and follicle arrest in the ovarian cortex of High A4 cows

Abedal-Majed¹,

Springman²,

Sutton³

et al. 2021

Biology of Reproduction

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A population of cows with excess androstenedione (A4; High A4) in follicular fluid, with follicular arrest, granulosa cell dysfunction, and a 17% reduction in calving rate was previously identified. We hypothesized that excess A4 in the ovarian microenvironment caused the follicular arrest in High A4 cows and Vascular Endothelial Growth Factor A (VEGFA) would rescue the High A4 phenotype. In trial 1, prior to culture, High A4 ovarian cortex (n = 9) had greater numbers of early stage follicles (primordial) and fewer later stage follicles compared to Controls (n = 11). Culture for 7 days did not relieve this follicular arrest; instead, High A4 ovarian cortex had increased indicators of inflammation, Anti-Mullerian Hormone (AMH) and A4 secretion compared to Controls. In trial 2, we tested if VEGFA isoforms could rescue the High A4 phenotype. High A4 (n = 5) and Control (n = 5) ovarian cortex was cultured with: 1) PBS; 2) VEGFA165 (50 ng/ml); 3) VEGFA165B (50 ng/ml); or 4) VEGFA165 + VEGFA165B (50 ng/ml each) for 7 days. Follicular progression increased with VEGFA165 in High A4 cows with greater early primary, primary, and secondary follicles than Controls. Similar to trial 1, High A4 ovarian cortex secreted greater concentrations of A4 and other steroids, and had greater indicators of inflammation compared to Controls. However, VEGFA165 rescued steroidogenesis, oxidative stress, and fibrosis. VEGFA165 and VEGFA165b both reduced IL-13, INFα, and INFβ secretion in High A4 cows to control levels. Thus, VEGFA165 may be a potential therapeutic to restore the ovarian steroidogenic microenvironment and promote folliculogenesis.

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Bovine Ovarian Cortex Tissue Culture

Cited by 5 publications

References 0 publications

New insights into ginsenoside Rg1 regulating the niche to inhibit age-induced germline stem cells depletion through targeting ECR/BMP signaling pathway in Drosophila

New insights into ginsenoside Rg1 regulating the niche to inhibit age-induced germline stem cells depletion through targeting ECR/BMP signaling pathway in Drosophila

Culture of vitrified bovine ovarian tissue on agarose gel inserts maintains follicle integrity

VEGFA165 can rescue excess steroid secretion, inflammatory markers, and follicle arrest in the ovarian cortex of High A4 cows

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