Novel dynamic culture system to support initiation of primordial follicle growth in prepubertal mouse ovaries

Winkler-Crepaz, Katharina; Nederegger, Verena; Ayuandari, Sarrah; Rosenfellner, Doris; Zervomanolakis, I.; Hofer, Susanne; Wildt, L.; Ziehr, Stephanie C.

doi:10.1016/j.fertnstert.2014.05.038

Cited by 13 publications

(7 citation statements)

References 37 publications

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“…In the present study, we established the visualization of follicle development using sliced ovarian tissue culture so that we could trace the follicle areas and track follicle development. Histological analysis using fixed ovarian tissues has been widely used to confirm possible follicle development in ovarian tissue culture [ 11 , 12 ]. However, using this method, it is impossible to obtain detailed information about which follicles, and when and how they are growing.…”

Section: Discussionmentioning

confidence: 99%

Follicle dynamics: visualization and analysis of follicle growth and maturation using murine ovarian tissue culture

Murase

Iwase

Komatsu

et al. 2017

J Assist Reprod Genet

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PurposeTo visualize and analyze follicle development in ovarian tissue culture using physiological concentrations of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in order to establish an ovarian tissue culture system that enables efficient in vitro growth of follicles.MethodsOvarian tissues from 4-week-old female ICR mice were sliced and cultured. Images of ovarian tissues in culture were obtained at 24-h or 30-min intervals by using a microscope. The area of each follicle observed in the ovarian tissue slices was tracked and analyzed in association with oocyte maturation.ResultsWe were able to track the development of each follicle using this culture system. Follicle growth was associated with oocyte maturation. Meiotically matured oocytes (MII) were obtained from 33% of all follicles investigated. Approximately, a quarter of follicles (24%) did not grow and resulted in atresia.ConclusionFollicle dynamics were successfully visualized and analyzed in murine ovarian tissue culture. We were able to obtain mature oocytes from the fully grown follicles in vitro. This culture system would be helpful for efficient in vitro culturing of ovarian tissues.Electronic supplementary materialThe online version of this article (10.1007/s10815-017-1073-5) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Follicle dynamics: visualization and analysis of follicle growth and maturation using murine ovarian tissue culture

Murase

Iwase

Komatsu

et al. 2017

J Assist Reprod Genet

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“…Intermediate follicles had both flattened and cuboidal granulosa cells. Only follicles with a visible nucleus in the oocyte were counted [21].…”

Section: Methodsmentioning

confidence: 99%

Randomized study to prove the quality of human ovarian tissue cryopreservation by xenotransplantation into mice

Ruan

Cui

et al. 2019

J Ovarian Res

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Purpose To study the quality of our human ovarian tissue cryopreservation technique as performed in the first official "International Fertility Protection Centre" in China in patients with certain cancer types using a mouse model, and to find the best site for tissue transplantation in the mouse. Methods Thirty-six BALB/C female nude mice were randomly divided into 3 groups, group 1: control group; group 2: ovariectomized group; group 3: ovarian tissue transplantation group. Seventy-two pieces obtained from six ovarian tissue samples from each of three cancer patients were transplanted into the ovarian bursa cavity (OBC), the subcutaneous thigh (TS) and the subcutaneous neck (NS) and removed after 1.5 and 2.5 months, respectively. Follicular growth rate (FGR), total follicle surviving rate (TFSR), tissue recovery rate (TRR), antral follicles (AF), follicle stimulating hormone (FSH), estradiol (E2) and anti-Mullerian hormone (AMH) levels were measured. Results No significant differences in FGR, OBC, NS ( p > 0.05); TFSR was 100% in OBC, NS and TS. No significant differences in TRR ( p > 0.05); AF were found only in OBC; TFSR was 100% after transplantation; significantly higher FGR in the 2.5 months compared to the 1.5 months-group ( p < 0.05). AMH- and E2-level in group 1 and 3 were significantly higher than in group 2 ( p < 0.05); in contrast, FSH was significantly lower. Conclusions After transplantation in the mice, the thawed ovarian tissue survived and follicles developed. The ovarian fossa site was the best site for transplantation. Our animal experiments can verify that our human ovarian tissue cryopreservation technique can preserve the quality of ovarian tissue. This is the essential precondition for successful re-transplantation into the patients after performing chemo/radiotherapy to protect ovarian function and fertility.

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“…In the case of intermediate stages between primordial and primary follicles, those follicles showing at least one cuboidal granulosa cell were classified as primary follicles. Only follicles with a visible nucleus in the oocyte were counted [47]. …”

Section: Methodsmentioning

confidence: 99%

Follicular growth after xenotransplantation of cryopreserved/thawed human ovarian tissue in SCID mice: dynamics and molecular aspects

Ayuandari

Winkler-Crepaz

Paulitsch

et al. 2016

J Assist Reprod Genet

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PurposeTo study the influence of xenotransplantation on follicular recruitment and growth in cryopreserved/thawed human ovarian tissue.MethodTwo 3-mm pieces of cryopreserved/thawed human ovarian tissue obtained from female cancer patients (n = 11) were xenotransplanted into a subcutaneous neck pouch of 6-week-old ovarectomized SCID mice (n = 33) for 4 (n = 18) and 12 (n = 15) weeks.ResultThirty-two out of 33 mice survived the entire observation periods. Graft recovery rate was 95.58 % (65 of 68 grafts). The percentages of primordial follicles after 4 weeks (P < 0.001) and 12 weeks (P = 0.009) of grafting were significantly lower in comparison to pregraft controls. The percentage of secondary follicle was significantly higher after 4 weeks of grafting (P = 0.018) and after 12 weeks (P = 0.001) of grafting in comparison to pregraft controls. Ki67 immunohistochemistry showed that proliferative follicles were significantly higher after 4 and 12 weeks of grafting compared to pregraft controls (P < 0.001). All follicles analyzed by TUNEL staining appeared healthy after xenotransplantation. The expression level of PTEN was reduced by 2.47-fold after 4 weeks of xenotransplantation, and this result was significant when 2−ΔCt were analyzed (P = 0.042).ConclusionThe higher proportion of growing follicles compared to resting follicles observed after xenotransplantation is most likely due to downregulation of PTEN gene expression followed by acceleration of follicular recruitment.Electronic supplementary materialThe online version of this article (doi:10.1007/s10815-016-0769-2) contains supplementary material, which is available to authorized users.

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Novel dynamic culture system to support initiation of primordial follicle growth in prepubertal mouse ovaries

Cited by 13 publications

References 37 publications

Follicle dynamics: visualization and analysis of follicle growth and maturation using murine ovarian tissue culture

Follicle dynamics: visualization and analysis of follicle growth and maturation using murine ovarian tissue culture

Randomized study to prove the quality of human ovarian tissue cryopreservation by xenotransplantation into mice

Follicular growth after xenotransplantation of cryopreserved/thawed human ovarian tissue in SCID mice: dynamics and molecular aspects

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