Effect of cryopreservation on the ability of granulosa cells to support in vitro development of oocytes derived from porcine early antral follicles

Ishiguro, Ai; Sakai, Hayato; Kansaku, Kazuki; Shirasuna, Koumei; Iwata, Hisataka

doi:10.1016/j.theriogenology.2019.11.038

Cited by 3 publications

(1 citation statement)

References 18 publications

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“…GCs in the wall of the follicle are in contact with the antrum and proliferate as the follicle grows. These cells are divided into two groups (mural granulosa cells and cumulus cells) based on their function and their distance from the oocyte [3]. Mural granulosa cells are responsible for the final maturation of follicles and are sensitive to estrogen and LH.…”

Section: Introductionmentioning

confidence: 99%

Optimized Primary Culture and Subculture of Granulosa Cells

Fallah¹,

Paktinat²,

Rasouli³

et al. 2022

PBR

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Background: Primordial follicle includes an oocyte surrounded by a layer of somatic cells called Granulosa Cells (GCs). GCs, also known as nurse cells, are an important protective element for the growth and survival of oocytes. Oocytes, which lack some of the metabolic processes, require granulosa cells for their development. Objectives: This manuscript was provided to explain the protocol of GCs primary culture extracted from NMRI mice ovaries. Methods: For choosing the optimum protocol, we used two methods with different culture mediums to obtain more GCs and expedite the process. Hematoxylin and Eosin (H&E) staining and flow cytometry were used to analyze the type of extracted cells from ovaries. Besides, we evaluated the effect of crocin and DPP as two common natural products in Iran on the proliferation of these cells via MTT assay. Results: Second protocol method and alpha-MEM culture medium were chosen based on the results. Our findings from HE staining and flow cytometry proved the percentage of cultured GCs in the flask. Further, MTT assessment demonstrated that crocin at high doses had a toxic effect on granulosa cells, whereas date palm pollen (DPP) stimulated them to proliferation. Conclusion: Modifying this protocol is for the improvement of proliferation, coherence, and quality of GCs in primary culture and subculture. Regarding the effect of these two natural products on granulosa cells, we can mention the bilateral effect of crocin and DPP enhancement in proliferation.

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Section: Introductionmentioning

confidence: 99%

Optimized Primary Culture and Subculture of Granulosa Cells

Fallah¹,

Paktinat²,

Rasouli³

et al. 2022

PBR

View full text Add to dashboard Cite

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Ovarian tissue features assessed in bovine fetuses after vitrification and xenotransplantation procedures

Moraes

Ronda

Campos

et al. 2021

Reproductive Biology

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Age-associated changes in miRNA profile of bovine follicular fluid

et al. 2022

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We examined age-associated changes in microRNA (miRNA) profiles in the follicular fluid (FF) of cows. The role of miR-19b, which is abundant in the FF of young cows, in in vitro growth of early antral follicles (EAFs)-derived oocytes was assessed. FF was collected from the antral follicles of young (20–40 months) and aged (>120 months) cows. The miRNA profiles were similar between the FF of both age groups, whereas the abundance of some miRNAs differed between these samples. The miRNA profiles in granulosa cells (GCs) and the spent culture medium of oocyte–GC complexes (OGCs) derived from EAFs were distinct. Some miRNA groups overlapped among the GCs, culture media, and FFs. miR-19b was highly abundant in the FF of young cows, GCs and culture medium. The supplementation of OGC culture medium with miR-19b increased the diameter, acetylation levels, and fertilisation ability of the oocytes. To assess whether miR-19b was functional in the GCs, a dual-luciferase assay, suppression of target protein, and RNA-sequencing of the GCs followed by functional annotation of the differentially expressed genes (DEGs) were conducted. Functional annotation of the DEGs suggested that miR-19b influences genes associated with FoxO signalling, endocytosis, and NR3C1 in GCs. These results suggest that in FFs, aging affects the abundance of miRNAs that have important roles in oocyte development.

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Effect of cryopreservation on the ability of granulosa cells to support in vitro development of oocytes derived from porcine early antral follicles

Cited by 3 publications

References 18 publications

Optimized Primary Culture and Subculture of Granulosa Cells

Optimized Primary Culture and Subculture of Granulosa Cells

Ovarian tissue features assessed in bovine fetuses after vitrification and xenotransplantation procedures

Age-associated changes in miRNA profile of bovine follicular fluid

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