scholarly journals ID3019 Female germline stem Cells: A source for application in reproductive and regenerative medicine

2017 ◽  
Vol 4 (S) ◽  
pp. 31
Author(s):  
Thuy Hong Bui
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Ovarian Function ◽  
Primordial Germ Cells ◽  
Postnatal Life ◽  
Proliferative Potential ◽  
Germline Stem Cells ◽  
Mammalian Female ◽  
Female Germline

Studies suggest a renewable source of eggs and stir more controversy, especially about the origin of female germline stem cells (FGSCs). It should be elucidated whether or not neo-oogenesis continues in the ovaries of mammalian female during postnatal life. Therefore, the establishment of FGSCs is very important for many applications. Here, using adult pig ovary, we isolate, identify, characterize FGSCs to elucidate their origin, then examined the proliferation, growth and differentiation of them. These cells were heterogeneous, depending on both of c-kit expression and cell size, and also express stem cell and germ cell markers. Importantly, we show clearly that the cells with the characteristics of early primordial germ cells are present in the adult pig ovary. Once FGSCs were established, they could be expanded in vitro for months without loss of the identifying markers and proliferative potential. Under appropriate conditions, the FGSCs differentiated into primordial oocyte-like cells and grow close to full-sized oocytes. These may assist in therapeutic strategies in human with their potential to make new oocytes and support ovarian function and fertility. Our results support the theory that the ovary contains a small number of undifferentiated cells with stem cell characteristics. These might remain in the postnatal and adult ovary and under certain conditions could resume mitosis, enter meiosis and give rise to oocytes. Given the existence of these FGSCs in mammalian ovaries and the depletion in ovarian reserve during female reproductive aging, one can hypothesize that such “neo-oogenesis” was present in ancestral forms, is still present in insects, some fish and mollusks, but has been lost in land vertebrates through evolution. FGSCs cannot proliferate in the ovary normally because of inhibitory factors, but under appropriate conditions, they can undergo proliferation and differentiation, and provide a potential mechanism for the self-renewal of germline stem cells.

scholarly journals ID: 1063 Isolation and characterization of female germline stem cell derived from porcine ovary

2017 ◽  
Vol 4 (S) ◽  
pp. 145
Author(s):  
Nguyen Huy Hoang ◽  
Nguyen Thi Bao Tran ◽  
Nguyen Van Thuan ◽  
Bui Hong Thuy
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
In Vitro Culture ◽  
Germline Stem Cell ◽  
Stem Cell Markers ◽  
Significant Finding ◽  
Germline Stem Cells ◽  
Isolation And Characterization ◽  
Female Germline

One of the most significant finding in stem cell area in the early 21st century is the founding of female germline stem cells (FGSCs). Establishment of FGSCs allowed new possibilities for the use of them in biotechnology and medicine. Hence, the purpose of this study was to establish, characterize the porcine female germline stem cells (pFGSCs) from porcine ovary. The result revealed the success in establishing pFGSCs from ovarian tissue. Most of the pFGSCs were round shape after in vitro culture, forming groups of cells that cluster around the ovarian cells colonies. Immunofluorescent analysis of pFGSCs showed that these cells expressed germ cell and stem cell markers such as: Vasa, Stella, c-kit and Oct4. After several weeks in in vitro culture, pFGSCs increased in number without the loss of proliferative potential. Our results suggested that pFGSCs isolated from adult mammalian ovary, under appropriate conditions, could undergo proliferation.

scholarly journals Comparison of different in vitro differentiation conditions for murine female germline stem cells

2018 ◽  
Vol 52 (1) ◽  
pp. e12530 ◽  
Author(s):  
Kang Zou ◽  
Jian Wang ◽  
Haiwei Bi ◽  
Yabin Zhang ◽  
Xueli Tian ◽  
...  
Keyword(s):  
Stem Cells ◽  
Germline Stem Cells ◽  
In Vitro Differentiation ◽  
Female Germline

scholarly journals Implications and Current Limitations of Oogenesis from Female Germline or Oogonial Stem Cells in Adult Mammalian Ovaries

Cells ◽  
2019 ◽  
Vol 8 (2) ◽  
pp. 93 ◽  
Author(s):  
Jessica Martin ◽  
Dori Woods ◽  
Jonathan Tilly
Keyword(s):  
Stem Cells ◽  
Adult Stem Cells ◽  
Ovarian Function ◽  
Mammalian Species ◽  
Large Body ◽  
The Body ◽  
Human Female ◽  
Female Germline

A now large body of evidence supports the existence of mitotically active germ cells in postnatal ovaries of diverse mammalian species, including humans. This opens the possibility that adult stem cells naturally committed to a germline fate could be leveraged for the production of female gametes outside of the body. The functional properties of these cells, referred to as female germline or oogonial stem cells (OSCs), in ovaries of women have recently been tested in various ways, including a very recent investigation of the differentiation capacity of human OSCs at a single cell level. The exciting insights gained from these experiments, coupled with other data derived from intraovarian transplantation and genetic tracing analyses in animal models that have established the capacity of OSCs to generate healthy eggs, embryos and offspring, should drive constructive discussions in this relatively new field to further exploring the value of these cells to the study, and potential management, of human female fertility. Here, we provide a brief history of the discovery and characterization of OSCs in mammals, as well as of the in-vivo significance of postnatal oogenesis to adult ovarian function. We then highlight several key observations made recently on the biology of OSCs, and integrate this information into a broader discussion of the potential value and limitations of these adult stem cells to achieving a greater understanding of human female gametogenesis in vivo and in vitro.

scholarly journals Fetal liver mesenchymal stem cells restore ovarian function in premature ovarian insufficiency by targeting MT1

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Boxian Huang ◽  
Chunfeng Qian ◽  
Chenyue Ding ◽  
Qingxia Meng ◽  
Qinyan Zou ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Stem Cell ◽  
Cell Therapy ◽  
Stem Cell Therapy ◽  
Ovarian Function ◽  
Ovarian Insufficiency ◽  
Apoptotic Genes

Abstract Background With the development of regenerative medicine and tissue engineering technology, almost all stem cell therapy is efficacious for the treatment of premature ovarian failure (POF) or premature ovarian insufficiency (POI) animal models, whereas little stem cell therapy has been practiced in clinical settings. The underlying molecular mechanism and safety of stem cell treatment in POI are not fully understood. In this study, we explored whether fetal mesenchymal stem cells (fMSCs) from the liver restore ovarian function and whether melatonin membrane receptor 1 (MT1) acts as a regulator for treating POI disease. Methods We designed an in vivo model (chemotherapy-induced ovary damage) and an in vitro model (human ovarian granulosa cells (hGCs)) to understand the efficacy and molecular cues of fMSC treatment of POI. Follicle development was observed by H&E staining. The concentration of sex hormones in serum (E2, AMH, and FSH) and the concentration of oxidative and antioxidative metabolites and the enzymes MDA, SOD, CAT, LDH, GR, and GPx were measured by ELISA. Flow cytometry (FACS) was employed to detect the percentages of ROS and proliferation rates. mRNA and protein expression of antiapoptotic genes (SURVIVIN and BCL2), apoptotic genes (CASPASE-3 and CASPASE-9), and MT1 and its downstream genes (JNK1, PCNA, AMPK) were tested by qPCR and western blotting. MT1 siRNA and related antagonists were used to assess the mechanism. Results fMSC treatment prevented cyclophosphamide (CTX)-induced follicle loss and recovered sex hormone levels. Additionally, fMSCs significantly decreased oxidative damage, increased oxidative protection, improved antiapoptotic effects, and inhibited apoptotic genes in vivo and in vitro. Furthermore, fMSCs also upregulated MT1, JNK1, PCNA, and AMPK at the mRNA and protein levels. With MT1 knockdown or antagonist treatment in normal hGCs, the protein expression of JNK1, PCNA, and AMPK and the percentage of proliferation were impaired. Conclusions fMSCs might play a crucial role in mediating follicular development in the POI mouse model and stimulating the activity of POI hGCs by targeting MT1.

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

2018 ◽  
Vol 46 (5) ◽  
pp. 2114-2126 ◽  
Author(s):  
Meng Wu ◽  
Jiaqiang Xiong ◽  
Lingwei Ma ◽  
Zhiyong Lu ◽  
Xian Qin ◽  
...  
Keyword(s):  
Stem Cells ◽  
Akt Pathway ◽  
Germline Stem Cells ◽  
Self Renewal ◽  
Differential Adhesion ◽  
Proliferation And Differentiation ◽  
Phosphoinositide 3 Kinase ◽  
Female Germline

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.

scholarly journals GAS5/miR-21 Axis as a Potential Target to Rescue ZCL-082-Induced Autophagy of Female Germline Stem Cells In Vitro

2019 ◽  
Vol 17 ◽  
pp. 436-447 ◽  
Author(s):  
Bo Li ◽  
Xiaopeng Hu ◽  
Yanzhou Yang ◽  
Mingyan Zhu ◽  
Jiong Zhang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Germline Stem Cells ◽  
Potential Target ◽  
Female Germline

scholarly journals C89 Induces Autophagy of Female Germline Stem Cells via Inhibition of the PI3K-Akt Pathway In Vitro

Cells ◽  
2019 ◽  
Vol 8 (6) ◽  
pp. 606 ◽  
Author(s):  
Xinyue Li ◽  
Xiaopeng Hu ◽  
Geng G. Tian ◽  
Ping Cheng ◽  
Zezhong Li ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Counting ◽  
Akt Pathway ◽  
Germline Stem Cell ◽  
Germline Stem Cells ◽  
Sequencing Data ◽  
Sequestosome 1 ◽  
Small Molecule Compound ◽  
Female Germline

Postnatal female germline stem cells (FGSCs) are a type of germline stem cell with self-renewal ability and the capacity of differentiation toward oocyte. The proliferation, differentiation, and apoptosis of FGSCs have been researched in recent years, but autophagy in FGSCs has not been explored. This study investigated the effects of the small-molecule compound 89 (C89) on FGSCs and the underlying molecular mechanism in vitro. Cytometry, Cell Counting Kit-8 (CCK8), and 5-ethynyl-2’-deoxyuridine (EdU) assay showed that the number, viability, and proliferation of FGSCs were significantly reduced in C89-treated groups (0.5, 1, and 2 µM) compared with controls. C89 had no impact on FGSC apoptosis or differentiation. However, C89 treatment induced the expression of light chain 3 beta II (LC3BII) and reduced the expression of sequestosome-1 (SQSTM1) in FGSCs, indicating that C89 induced FGSC autophagy. To investigate the mechanism of C89-induced FGSC autophagy, RNA-seq technology was used to compare the transcriptome differences between C89-treated FGSCs and controls. Bioinformatics analysis of the sequencing data indicated a potential involvement of the phosphatidylinositol 3 kinase and kinase Akt (PI3K-Akt) pathway in the effects of C89′s induction of autophagy in FGSCs. Western blot confirmed that levels of p-PI3K and p-Akt were significantly reduced in the C89- or LY294002 (PI3K inhibitor)-treated groups compared with controls. Moreover, we found cooperative functions of C89 and LY294002 in inducing FGSC autophagy through suppressing the PI3K-Akt pathway. Taken together, this research demonstrates that C89 can reduce the number, viability, and proliferation of FGSCs by inducing autophagy. Furthermore, C89 induced FGSC autophagy by inhibiting the activity of PI3K and Akt. The PI3K-Akt pathway may be a target to regulate FGSC proliferation and death.

Lentiviral vector transfection of chicken embryo primordial germ cells in vitro

2008 ◽  
Vol 5 (1) ◽  
pp. 1-5
Author(s):  
Ding Hong-Mei ◽  
Xu Shi-Yong ◽  
Shao Gen-Bao ◽  
Sun Yan ◽  
Wang Meng ◽  
...  
Keyword(s):  
Stem Cells ◽  
Gene Transfer ◽  
Germ Cells ◽  
Lentiviral Vector ◽  
Transfection Efficiency ◽  
Primordial Germ Cells ◽  
Embryonic Stem ◽  
Germline Stem Cells ◽  
Transfer Vectors

AbstractLentiviruses as gene transfer vectors have been used successfully to transfect mammal embryonic stem cells and germline stem cells, but this has not been attempted in avian primordial germ cells (PGCs). PGCs were isolated from the gonads of Isa-brown chicken embryos at stage 28 and co-cultured with gonadal stroma cells. A lentiviral vector pLenti-CMV-EGFP was constructed and the virus harvested by cotransfecting 293FT cells with the vector and packaging plasmids. Concentrated lentiviruses were used to transfect chicken PGCs, the transfection efficiency was up to 24.19%.

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

PLoS ONE ◽  
2015 ◽  
Vol 10 (10) ◽  
pp. e0139824 ◽  
Author(s):  
Jiaqiang Xiong ◽  
Zhiyong Lu ◽  
Meng Wu ◽  
Jinjin Zhang ◽  
Jing Cheng ◽  
...  
Keyword(s):  
Stem Cells ◽  
Ovarian Function ◽  
Germline Stem Cells ◽  
Female Germline

scholarly journals Effects of shikonin on the development of ovarian follicles and female germline stem cells

2021 ◽  
Vol 49 (7) ◽  
pp. 030006052110294
Author(s):  
Shu-Xin Ma ◽  
Li-Bo Tang ◽  
Zhi-Hang Chen ◽  
Min-Li Wei ◽  
Zi-Juan Tang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Ovarian Follicles ◽  
Germline Stem Cell ◽  
Stem Cell Markers ◽  
Germline Stem Cells ◽  
Primordial Follicles ◽  
Cell Markers ◽  
Female Germline ◽  
Ovarian Index

Objective To investigate the effects and potential mechanism of action of shikonin (SHK) on the development of ovarian follicles and female germline stem cells (FGSCs). Methods Female Kunming adult mice were administered SHK (0, 20 and 50 mg/kg) by oral gavage. Cultures of FGSCs were treated with SHK 32 μmol/l for 24 h. The ovarian index in mouse ovaries was calculated. Numbers of primordial, primary and atretic follicles were counted. Germline stem cell markers and apoptosis were examined. Levels of glutathione (GSH), superoxide dismutase (SOD) and reactive oxygen species (ROS) were measured. Results Both doses of SHK significantly decreased the ovarian index, the numbers of primordial follicles, primary follicles and antral follicles in mice. SHK significantly increased the numbers of atretic follicles and atretic corpora lutea. SHK promoted apoptosis in vivo and in vitro. SHK significantly decreased the levels of the germline stem cell markers. SHK significantly lowered GSH levels and the activity of SOD in the peripheral blood from mice, whereas SHK significantly elevated cellular ROS content in FGSCs. Conclusions These current results suggested that follicular development and FGSCs were suppressed by SHK through the induction of apoptosis and oxidative stress might be involved in this pathological process.

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