scholarly journals Expression of Pluripotency and Oocyte-Related Genes in Single Putative Stem Cells from Human Adult Ovarian Surface Epithelium CulturedIn Vitroin the Presence of Follicular Fluid

2013 ◽  
Vol 2013 ◽  
pp. 1-18 ◽  
Author(s):  
Irma Virant-Klun ◽  
Thomas Skutella ◽  
Mikael Kubista ◽  
Andrej Vogler ◽  
Jasna Sinkovec ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Cultures ◽  
Follicular Fluid ◽  
Premature Ovarian Failure ◽  
Ovarian Surface Epithelium ◽  
Ovarian Failure ◽  
Reproductive Hormones ◽  
Surface Epithelium ◽  
Oocyte Growth ◽  
Ovarian Surface

The aim of this study was to trigger the expression of genes related to oocytes in putative ovarian stem cells scraped from the ovarian surface epithelium of women with premature ovarian failure and culturedin vitroin the presence of follicular fluid, rich in substances for oocyte growth and maturation. Ovarian surface epithelium was scraped and cell cultures were set up by scrapings in five women with nonfunctional ovaries and with no naturally present mature follicles or oocytes. In the presence of donated follicular fluid putative stem cells grew and developed into primitive oocyte-like cells. A detailed single-cell gene expression profiling was performed to elucidate their genetic status in comparison to human embryonic stem cells, oocytes, and somatic fibroblasts. The ovarian cell cultures depleted/converted reproductive hormones from the culture medium. Estradiol alone or together with other substances may be involved in development of these primitive oocyte-like cells. The majority of primitive oocyte-like cells was mononuclear and expressed several genes related to pluripotency and oocytes, including genes related to meiosis, although they did not express some important oocyte-specific genes. Our work reveals the presence of putative stem cells in the ovarian surface epithelium of women with premature ovarian failure.

scholarly journals Isolation of Small SSEA-4-Positive Putative Stem Cells from the Ovarian Surface Epithelium of Adult Human Ovaries by Two Different Methods

2013 ◽  
Vol 2013 ◽  
pp. 1-15 ◽  
Author(s):  
Irma Virant-Klun ◽  
Thomas Skutella ◽  
Matjaz Hren ◽  
Kristina Gruden ◽  
Branko Cvjeticanin ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Sorting ◽  
Primordial Germ Cell ◽  
Ovarian Surface Epithelium ◽  
Reproductive Age ◽  
Small Cells ◽  
Surface Epithelium ◽  
Oocyte Growth ◽  
Adult Human ◽  
Ovarian Surface

The adult ovarian surface epithelium has already been proposed as a source of stem cells and germinal cells in the literature, therefore it has been termed the “germinal epithelium”. At present more studies have confirmed the presence of stem cells expressing markers of pluripotency in adult mammalian ovaries, including humans. The aim of this study was to isolate a population of stem cells, based on the expression of pluripotency-related stage-specific embryonic antigen-4 (SSEA-4) from adult human ovarian surface epithelium by two different methods: magnetic-activated cell sorting and fluorescence-activated cell sorting. Both methods made it possible to isolate a similar, relatively homogenous population of small, SSEA-4-positive cells with diameters of up to 4 μm from the suspension of cells retrieved by brushing of the ovarian cortex biopsies in reproductive-age and postmenopausal women and in women with premature ovarian failure. The immunocytochemistry and genetic analyses revealed that these small cells—putative stem cells—expressed some primordial germ cell and pluripotency-related markers and might be related to thein vitrodevelopment of oocyte-like cells expressing some oocyte-specific transcription factors in the presence of donated follicular fluid with substances important for oocyte growth and development. The stemness of these cells needs to be further researched.

Isolation and Characterization of Mesenchymal Stem Cells from Canine Ovarian Surface Epithelium

2021 ◽  
Author(s):  
Ajeet Kumar Jha ◽  
Anirban Mandal ◽  
Kalyani Ray ◽  
Shyamal Kanti Guha
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Stromal Vascular Fraction ◽  
Ovarian Surface Epithelium ◽  
Surface Epithelium ◽  
High Plasticity ◽  
Rt Pcr ◽  
Differentiation Potential ◽  
Ovarian Surface ◽  
Isolation And Characterization

Background: Few studies have confirmed the presence of ovarian tissue stem cells indicating the capacity for differentiation. Based on this fact, it was hypothesized that mesenchymal stem cells (MSC) were found in ovarian surface epithelium (OSE) of canines that could easily be isolated. Methods: Both left and right ovaries were minced and digested using collagenase to obtain a stromal vascular fraction (SVF). MSCs were characterized using RT-PCR. To ascertain the trilineage differentiation potential, MSCs were stained with respective stain for osteocytes, chondrocytes and adipocytes. Result: We observed elongated, spindle-shaped and fibroblast like appearance of cells after 72 h of initial culture. Expression of MSC specific surface markers were observed through RT-PCR. Using Stem Pro® differentiation medium, OSE were differentiated into osteogenic, chondrogenic and adipogenic lineages and were found to be potential source for isolation, characterization and differentiation of MSCs. Canine (OSE) is easily accessible, multipotent and has high plasticity, holding promise for applications in regenerative medicine.

scholarly journals The Expression of Markers Related to Ovarian Germline Stem Cells in the Mouse Ovarian Surface Epithelium and the Correlation with Notch Signaling Pathway

2015 ◽  
Vol 37 (6) ◽  
pp. 2311-2322 ◽  
Author(s):  
Zezheng Pan ◽  
Mengli Sun ◽  
Jia Li ◽  
Fangyue Zhou ◽  
Xia Liang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Notch Signaling ◽  
Signaling Pathway ◽  
Ovarian Surface Epithelium ◽  
Notch Signaling Pathway ◽  
Surface Epithelium ◽  
Stem Cell Markers ◽  
Germline Stem Cells ◽  
Ovarian Surface

Background/Aims: Ovarian germline stem cells (OGSCs) have been shown to mainly exist in the ovarian surface epithelium (OSE), but the activity changes of germline stem cells during different reproductive stages and the potential regulatory signaling pathway are still unknown. The Notch signaling pathway plays a key role in cell development, primordial follicles and stem cell proliferation. However, whether it plays a role in the proliferation of OGSCs is unknown. Here, we analyzed the activity changes of germline stem cells and the correlation between germline stem cells and the Notch signaling pathway. Methods: The expression of germline stem cell markers Mvh, Ooc4 and the Notch molecules Notch1, Hes1, and Hes5 were detected during 3 days (3d), and 2, 12, 20 months (2m, 12m, 20m) mouse ovarian surface epithelium samples. DAPT, a specific inhibitor of the Notch pathway, was used to observe the influence of Notch signaling in the germline stem cells. Results: The results showed that the levels of MVH and OCT4 decreased substantially with reproductive age in ovarian surface epithelium, and the same tendency was detected in the Notch signaling molecules Notch1, Hes1 and Hes5. Dual-IF results showed that the germline stem cell markers were co-expressed with Notch molecules in the ovarian surface epithelium. While, the expression of MVH and OCT4 were reduced when the ovaries were treated with DAPT and the levels were attenuated with increasing dose of DAPT. Conclusion: Taken together, our results indicate that the viability of OGSCs decreased with the age of the mouse ovaries, and the activity of OGSCs in the ovarian surface epithelium may be related to the Notch signaling pathway.

333 GENERATION OF OOCYTE-LIKE STRUCTURE FROM OVARIAN SURFACE EPITHELIAL STEM CELLS OF GOAT

2015 ◽  
Vol 27 (1) ◽  
pp. 255 ◽  
Author(s):  
S. Fatima ◽  
V. Sharma ◽  
S. Saini ◽  
S. Saugandhika ◽  
H. N. Malik ◽  
...  
Keyword(s):  
Stem Cells ◽  
Endangered Species ◽  
Epithelial Cells ◽  
Ovarian Surface Epithelium ◽  
Surface Epithelium ◽  
Epithelial Stem Cells ◽  
Rt Pcr ◽  
Ovarian Surface ◽  
Ovarian Surface Epithelial

Stem cells have potential for therapeutic application. Continuous repair of ovarian surface epithelium following folliculogenesis and ovarian carcinoma suggests the presence of stem cells in ovarian epithelial cells. In vitro gametogenesis in livestock will result in large numbers of oocytes production from a single ovary, resulting in faster multiplication of superior germplasm of livestock species, treatment of infertile animals, and conservation of endangered species. The present study was conducted with the objective of in vitro differentiation of putative ovarian surface epithelial stem cells into oocyte-like structures in goat model. Ovary samples of 1- to 2-year-old goats were collected from slaughterhouse. The surface of the ovary was gently scraped using sterile blunt scraper to isolate ovarian surface epithelial stem cells. These scraped cells were cultured in DMEM/F12 supplemented with 20% FBS for 3 weeks in 5% CO2 at 37°C with maximum humidity. The cultured stem cells were characterised for stemness by RT-PCR and immunostaining for Oct4, Sox2, and Nanog genes after 3 weeks. These putative stem cells were in vitro differentiated spontaneously to oocyte-like structures in DMEM/F12 medium and characterised for premeiotic markers by RT-PCR and immunostaining for VASA, DAZL, and STELLA genes. Results of this study provide evidence for the presence of putative stem cells with pluripotent characteristics in the ovarian surface epithelium. The cultured cells were found to be round in shape, with a high nucleus to cytoplasm ratio under inverted microscope, and found positive for stem cell markers of Oct4, Sox2, and Nanog genes. A total of 66 oocyte-like structures were produced from 12 ovaries. These oocyte-like structures were nearly similar to oocytes produced in vivo, both morphologically and in molecular gene expression. The oocyte-like structures were also found positive for premeiotic markers of VASA, DAZL, and STELLA genes by RT-PCR and immunostaining. From this study, we concluded that the ovarian surface epithelial cells have putative stem cells which can be in vitro differentiated into oocyte-like structures in goat. These oocyte-like structures need further characterisation of their surface membrane, more molecular markers, and following their developmental potential. These oocytes can help for multiplication of elite germplasm, curing infertile animals, and saving endangered species.

Putative stem cells with an embryonic character isolated from the ovarian surface epithelium of women with no naturally present follicles and oocytes

2008 ◽  
Vol 76 (8) ◽  
pp. 843-856 ◽  
Author(s):  
Irma Virant-Klun ◽  
Nicolas Zech ◽  
Primož Rožman ◽  
Andrej Vogler ◽  
Branko Cvjetičanin ◽  
...  
Keyword(s):  
Stem Cells ◽  
Ovarian Surface Epithelium ◽  
Surface Epithelium ◽  
Ovarian Surface

scholarly journals Ovarian Surface Epithelium in Patients with Severe Ovarian Infertility: A Potential Source of Cells Expressing Markers of Pluripotent/Multipotent Stem Cells

2011 ◽  
Vol 2011 ◽  
pp. 1-12 ◽  
Author(s):  
Irma Virant-Klun ◽  
Thomas Skutella ◽  
Martin Stimpfel ◽  
Jasna Sinkovec
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Unknown Origin ◽  
Ovarian Surface Epithelium ◽  
Surface Epithelium ◽  
Stem Cell Line ◽  
Multipotent Stem Cells ◽  
Ovarian Surface ◽  
Potential Source ◽  
Shaped Cell

The aim of this study was to confirm the presence of stem cells in the ovarian surface epithelium of patients with premature ovarian failure and no mature follicles and oocytes. In these patients, small round cells of unknown origin expressing SOX-2 marker of pluripotency were observed among the epithelial cells just after the ovarian surface epithelium scraping. These cells were an integral part of the ovarian surface epithelium. When the scraped cells were cultured in a medium with added follicular fluid to provide some ovarian niche, primitive oocyte-like cells and typical round-shaped cell clusters positively stained on alkaline phosphatase, and markers of pluripotency, such as SOX-2 and SSEA-4, were developed. These markers were expressed early and also later in the culture. Single oocyte-like cells expressed genesOCT4A, SOX-2, NANOG, NANOS, STELLA, CD9, LIN28, KLF4, GDF3,andMYC, characteristic for pluripotent stem cells. The results of this study confirmed the presence of putative stem cells in the ovarian surface epithelium of these patients and provided some basis to create a stem cell line in the future.

scholarly journals Effects of coenzyme Q10 on ovarian surface epithelium-derived ovarian stem cells and ovarian function in a 4-vinylcyclohexene diepoxide-induced murine model of ovarian failure

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Hyun Joo Lee ◽  
Min Jung Park ◽  
Bo Sun Joo ◽  
Jong Kil Joo ◽  
Yeon Hee Kim ◽  
...  
Keyword(s):  
Stem Cells ◽  
Coenzyme Q10 ◽  
Ovarian Function ◽  
Ovarian Surface Epithelium ◽  
Ovarian Failure ◽  
Development Rate ◽  
Surface Epithelium ◽  
Vinylcyclohexene Diepoxide ◽  
Ovarian Aging ◽  
Ovarian Stem Cells

Abstract Background Several studies have shown that coenzyme Q10 (CoQ10) can rescue ovarian aging and that ovarian surface epithelium (OSE)-derived ovarian stem cells (OSCs) are useful for treating infertility due to ovarian aging. However, few studies have examined the effect of CoQ10 on OSCs. This study was aimed to investigate whether CoQ10 activates OSCs and recovers ovarian function in a 4-vinylcyclohexene diepoxide (VCD)-induced mouse model of ovarian failure. Methods Forty female C57BL/6 mice aged 6 weeks were randomly divided into four groups (n = 10/group): a control group administered saline orally, a CoQ10 group administered 150 mg/kg/day of CoQ10 orally in 1 mL of saline daily for 14 days, a VCD group administered 160 mg/kg/day of VCD i.p. in 2.5 mL of saline/kg for 5 days, and a VCD + CoQ10 group administered VCD i.p. for 5 days injection and CoQ10 (150 mg/kg/day) orally for 14 days. After treatment, follicle counts were evaluated by hematoxylin and eosin (H&E) staining, and ovarian mRNA expressions of Bmp-15, Gdf-9, and c-Kit were examined by quantitative real-time PCR. Serum FSH, AMH, and ROS levels were also measured. Oocyte-like structure counts and the expressions of Oct-4 and MVH were also evaluated after culturing OSE for 3 weeks. In a second experiment, 32 female mice were administered CoQ10 as described above, induced to superovulate using PMSG and hCG, and mated. Numbers of zygotes and embryo development rate were examined. Results Postcultured OSE showed significant increases in the numbers of oocyte-like structure and that the expression of Oct-4 and MVH were higher in the VCD + CoQ10 group than in the VCD group (p < 0.05). Numbers of surviving follicles from primordial to antral follicles, numbers of zygotes retrieved and embryo development rate to blastocyst were significantly greater in the VCD + CoQ10 group than in the VCD group (p < 0.01). Serum AMH level and ovarian expressions of Bmp-15, Gdf-9 and c-Kit were also significantly greater in the VCD + CoQ10 group than in the VCD group (p < 0.05). In contrast, serum ROS level was significantly lower in the VCD + CoQ10 group than in the VCD group (p < 0.05). Conclusion This study shows that CoQ10 stimulates the differentiation of OSE-derived OSCs and confirms that CoQ10 can reduce ROS levels and improve ovarian function and oocyte quality in mice with VCD-induced ovarian failure.

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