Cell Therapy for Chemically Induced Ovarian Failure in Mice

Terraciano, Paula Barros; Garcez, Tuane Nerissa Alves; Ayres, Laura Silveira; Durli, Isabel Cirne Lima de Oliveira; Baggio, Melchiani; Kuhl, Cristiana Palma; Laurino, Claudia Cilene Fernandes Correia; Passos, Eduardo Pandolfi; Paz, Ana Helena da Rosa; Cirne-Lima, Elizabeth Obino

doi:10.1155/2014/720753

Cited by 50 publications

(34 citation statements)

References 31 publications

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“…first confirmed the presence of FGSCs in mouse ovaries, many other researchers have gradually confirmed the existence of FGSCs in other mammalian species [34-36]. Furthermore, several research groups have extended the application of FGSCs in different fields, such as in vitro differentiation into oocytes [37], generation of transgenic mice [38], mechanism research [39, 40] and cell therapy [41, 42]. Although the supporters are in favour of the existence of FGSCs, there are still a number of opponents.…”

Section: Discussionmentioning

confidence: 99%

Enrichment of Female Germline Stem Cells from Mouse Ovaries Using the Differential Adhesion Method

Xiong

et al. 2018

Cell Physiol Biochem

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Background/Aims: The isolation and establishment of female germline stem cells (FGSCs) is controversial because of questions regarding the reliability and stability of the isolation method using antibody targeting mouse vasa homologue (MVH), and the molecular mechanism of FGSCs self-renewal remains unclear. Thus, there needs to be a simple and reliable method for sorting FGSCs to study them. Methods: We applied the differential adhesion method to enrich FGSCs (DA-FGSCs) from mouse ovaries. Through four rounds of purification and 7-9 subsequent passages, DA-FGSC lines were established. In addition, we assessed the role of the phosphoinositide-3 kinase (PI3K)-AKT pathway in regulating FGSC self-renewal. Results: The obtained DA-FGSCs spontaneously differentiated into oocyte-like cells in vitro and formed functional eggs in vivo that were fertilized and produced healthy offspring. AKT was rapidly phosphorylated when the proliferation rate of FGSCs increased after 10 passages, and the addition of a chemical PI3K inhibitor prevented FGSCs self-renewal. Furthermore, over-expression of AKT-induced proliferation and differentiation of FGSCs, c-Myc, Oct-4 and Gdf-9 levels were increased. Conclusions: The differential adhesion method provides a more feasible approach and is an easier procedure to establish FGSC lines than traditional methods. The AKT pathway plays an important role in regulation of the proliferation and maintenance of FGSCs. These findings could help promote stem cell studies and provide a better understanding of causes of ovarian infertility, thereby providing potential treatments for infertility.

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Section: Discussionmentioning

confidence: 99%

Enrichment of Female Germline Stem Cells from Mouse Ovaries Using the Differential Adhesion Method

Xiong

et al. 2018

Cell Physiol Biochem

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“…Nonetheless, a number of studies have verified that FGSCs exist in adult mammalian ovaries and could be isolated and cultured in vitro for more than 15 months [ 8 , 13 , 17 , 18 ]. Recently, a stem cell-based treatment for the protection of ovarian function and oocyte formation has been proposed as a future clinical therapy to treat infertility in female animals [ 9 , 12 , 19 , 20 ]. However, whether the ovaries of women treated with CTx also contain FGSCs and whether the aforementioned FGSCs could be used to protect gonadal function and fertility remains unknown.…”

Section: Discussionmentioning

confidence: 99%

Intraovarian Transplantation of Female Germline Stem Cells Rescue Ovarian Function in Chemotherapy-Injured Ovaries

Xiong

et al. 2015

PLoS ONE

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Early menopause and infertility often occur in female cancer patients after chemotherapy (CTx). For these patients, oocyte/embryo cryopreservation or ovarian tissue cryopreservation is the current modality for fertility preservation. However, the above methods are limited in the long-term protection of ovarian function, especially for fertility preservation (very few females with cancer have achieved pregnancy with cryopreserved ovarian tissue or eggs until now). In addition, the above methods are subject to their scope (females with no husband or prepubertal females with no mature oocytes). Thus, many females who suffer from cancers would not adopt the above methods pre- and post-CTx due to their uncertainty, safety and cost-effectiveness. Therefore, millions of women have achieved long-term survival after thorough CTx treatment and have desired to rescue their ovarian function and fertility with economic, durable and reliable methods. Recently, some studies showed that mice with infertility caused by CTx can produce normal offspring through intraovarian injection of exogenous female germline stem cells (FGSCs). Though exogenous FGSC can be derived from mice without immune rejection in the same strain, it is difficult to obtain human female germline stem cells (hFGSCs), and immune rejection could occur between different individuals. In this study, infertility in mice was caused by CTx, and the ability of FGSCs to restore ovarian function or even produce offspring was assessed. We had successfully isolated and purified the FGSCs from adult female mice two weeks after CTx. After infection with GFP-carrying virus, the FGSCs were transplanted into ovaries of mice with infertility caused by CTx. Finally, ovarian function was restored and the recipients produced offspring long-term. These findings showed that mice with CTx possessed FGSCs, restoring ovarian function and avoiding immune rejection from exogenous germline stem cells.

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“…Different doses of lentiviral particles were injected into the ovarian envelope under the microscope in vitro ; in the control group, lentivirus was replaced with the same volume of 0.9% normal saline solution (Fig. 1) [25]. After injections, the ovaries were placed back into the abdomen and incisions were disinfected and sutured.…”

Section: Methodsmentioning

confidence: 99%

The Hippo Signaling Pathway Regulates Ovarian Function via the Proliferation of Ovarian Germline Stem Cells

Zheng

et al. 2017

Cell Physiol Biochem

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Objective: To improve the separation, identification and cultivation of ovarian germline stem cells (OGSCs), to clarify the relationship between the Hippo signaling pathway effector YAP1 and the proliferation and differentiation of OGSCs in vitro and to identify the major contribution of Hippo signaling to ovarian function. Methods: Two-step enzymatic separation processes and magnetic separation were used to isolate and identify OGSCs by determining the expression of Mvh, Oct4, Nanog, Fragilis and Stella markers. Then, YAP1, as the main effector molecule in the Hippo signaling pathway, was chosen as the target gene of the study. Lentivirus containing overexpressed YAP1 or a YAP1-targeted shRNA was transduced into OGSCs. The effects of modulating the Hippo signaling pathway on the proliferation, differentiation, reproduction and endocrine function of ovaries were observed by microinjecting the lentiviral vectors with overexpressed YAP1 or YAP1 shRNA into infertile mouse models or natural mice of reproductive age. Results: (1) The specific expression of Mvh, Oct4, Nanog, Fragilis and Stella markers was observed in isolated stem cells. Thus, the isolated cells were preliminarily identified as OGSCs. (2) The co-expression of LATS2, MST1, YAP1 and MVH was observed in isolated OGSCs. Mvh and Oct4 expression levels were significantly increased in OGSCs overexpressing YAP1 compared to GFP controls. Consistently, Mvh and Oct4 levels were significantly decreased in cells expressing YAP1-targeted shRNA. (3) After 14-75 days of YAP1 overexpression in infertile mouse models, we detected follicle regeneration in ovaries, the activation of primordial follicles and increased birth rate, accompanied by increasing levels of E2 and FSH. (4) However, we detected decreasing follicles in ovaries, lower birth rate, and decreasing E2 and FSH in serum from healthy mice of reproductive age following YAP1 shRNA expression. Conclusion: Methods for the isolation, identification and culture of OGSCs were successfully established. Further results indicate that isolated OGSCs can specifically recognize Hippo signaling molecules and that manipulation of YAP1 expression can be used to regulate the proliferation and differentiation of OGSCs, as well as ovarian function in mice. This study suggests that the Hippo signaling pathway may represent a new molecular target for the regulation of mouse ovarian functional remodeling.

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Cell Therapy for Chemically Induced Ovarian Failure in Mice

Cited by 50 publications

References 31 publications

Enrichment of Female Germline Stem Cells from Mouse Ovaries Using the Differential Adhesion Method

Enrichment of Female Germline Stem Cells from Mouse Ovaries Using the Differential Adhesion Method

Intraovarian Transplantation of Female Germline Stem Cells Rescue Ovarian Function in Chemotherapy-Injured Ovaries

The Hippo Signaling Pathway Regulates Ovarian Function via the Proliferation of Ovarian Germline Stem Cells

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