Establishment of an <i>in vitro</i> culture model of theca cells from hierarchical follicles in ducks

Gan, Xiang; chen, da; Deng, Yan; Yuan, Junsong; Kang, Bo; Qiu, Jiamin; Sun, Wenqiang; Han, Chunchun; Hu, Jiwei; Li, Liang; Wang, Jiwen

doi:10.1042/bsr20160491

Cited by 17 publications

(20 citation statements)

References 41 publications

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“…Thereafter, the cells came into the plateau stage within the incubation of 120–144 h, followed by the senescence phase ( Figure 4 ). These results altogether suggested that the three populations of geese GCs showed both similar and different growth characteristics during in vitro culture, which was to a larger extent consistent with the phenomenon seen in other cell types [ 31 ]. Additionally, the identification of isolated geese GCs were verified by IF assay with the specific FSHR primary antibody, showing that no other cell contaminations were present in all three cohorts of GCs ( Figure 5 ).…”

Section: Discussionsupporting

confidence: 58%

Comparison of growth characteristics of in vitro cultured granulosa cells from geese follicles at different developmental stages

et al. 2018

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Granulosa cells (GCs) are essential components of follicles and are involved in regulating the process of follicles development. However, comparative studies on GCs isolated from different staged follicles have not been conducted in goose. The aim of the present study was to identify the growth characteristics of goose GCs from pre-hierarchical (6–10 mm) and hierarchical (F4–F2, F1) follicles. Our results showed that the three cohorts of cells had different tolerance to collagenase and had noticeable morphological differences. The F1 granulosa layers were fully digested by 0.1% collagenase, while higher concentration (0.3%) was used for both F4–F2 and pre-hierarchical granulosa layers. In the state of suspension, the diameter of F1 individual cell was larger than the other two cohorts. However, after adhering to the culture plate, cells of F1 just had changes in the diameter accompanied by small bright spots, while both pre-hierarchical and F4–F2 GCs proliferated rapidly with spreading and irregularly shaped voids. Furthermore, all attached cells could be stained by the follicle-stimulating hormone receptor antibody. Analyses of both growth curve and the mRNA expression profiles of genes related to cellular proliferation, apoptosis, and steroidogenesis suggested that three cohorts of in vitro cultured GCs had different physiological viability and functions. Taken together, the present study not only revealed differences of the growth characteristics among three cohorts of goose GCs from pre-hierarchical, F4–F2 and F1 follicles, but also optimized the in vitro culture system of geese different staged GCs.

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Section: Discussionsupporting

confidence: 58%

Comparison of growth characteristics of in vitro cultured granulosa cells from geese follicles at different developmental stages

et al. 2018

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“…It has been demonstrated that the mammalian ovary is mainly composed of several types of cells, including germ cells, surface epithelium cells, granulosa cells, theca, and stroma cells . Moreover, the ovarian cells, including somatic cells and germ cells, have been widely identified and isolated for in vitro culture . Especially, various methods were recently adopted to isolate and characterize the fGSCs in mammals, yet such studies in aquatic species are still scarce .…”

Section: Discussionmentioning

confidence: 99%

Isolation and in vitro culture of ovarian stem cells in Chinese soft‐shell turtle (Pelodiscus sinensis)

Zhu

et al. 2018

J of Cellular Biochemistry

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Gonadal cell lines provide valuable tools for studying gametogenesis, sex differentiation, and manipulating germ cells in reproductive biology. Female germline stem cells have been characterized and isolated from ovaries of mammalian species, including mice and human, but there has been very few studies on female germline stem cells in reptiles. Here, we described an ovarian stem cell-like line isolated and cultured from the Chinese soft-shell turtle (Pelodiscus sinensis), designated as PSO1. The cells showed high alkaline phosphatase activity with a normal diploid karyotype. As shown by reverse transcription-polymerase chain reaction, the cells were positive for the expression of germ cell-specific genes, vasa and dazl, as well as a stem cell marker, nanog, but negative for the expression of the folliculogenesis-specific gene, figla. Likewise, through fluorescent immunostaining analyses, both the Dazl and Vasa proteins were detected abundantly in the cytoplasm of perinuclear region, whereas Nanog and PCNA were dominantly observed in the nuclei in PSO1 cells. Moreover, PSO1 cells transfected with pCS2:h2b-egfp could properly express the fusion protein in the nuclei. Taken together, the findings suggested that the germline stem cells exist in the ovary of juvenile Chinese soft-shell turtle and these cells can be isolated for a long-term in vitro culture under experimental conditions. This study has provided a valuable basis for further investigations on the molecular mechanisms whereby the germline stem cells develop and differentiate into gametes in turtles. Also, it has paved the way for studies on oogenesis in turtles, even in the other reptiles.

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“…23 Therefore, during the initial passage of theca cells by trypsin-EDTA, the residual GCs could be removed. While responding this question, it should be noted that because GCs are separated from the theca cells by the base membrane, it seems that only theca cells are isolated for culture by puncturing of follicles and removing the contents within it by spatula.…”

Section: Discussionmentioning

confidence: 99%

Establishment and characterization of human theca stem cells and their differentiation into theca progenitor cells

Dalman

Totonchi

Valojerdi

2018

J of Cellular Biochemistry

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In this study, we have characterized the human theca stem cells (hTSCs) and their differentiation into human theca progenitor cells (hTPCs). hTSCs were isolated from the theca layer of small antral follicles (3-5 mm in size). Alkaline phosphatase activity, cell cycle status, and cell surface markers were evaluated in hTSCs. The differentiation potential of these cells was investigated via differentiation of hTSCs into adipocyte-, osteocyte-, and chondrocyte-like cells. The cells also differentiated into hTPCs. The hTSCs were morphologically similar to human fibroblast cells (hFCs). Some of the cells were positive for alkaline phosphatase activity. The expression of OCT4 in hTSCs was significantly higher than that of human bone marrow mesenchymal stem cells (hBMSCs) and hFCs. To determine the type of OCT4 (isoform A or B), RT PCR was performed. The data showed that OCT-4A was expressed in hBMSCs and hTSCs but immunofluorescence analyses using the OCT-4A-specific and OCT4 antibodies did not show OCT-4A protein. In addition, cell cycle status showed that the number of hTSCs in the S phase was significantly higher than that of hFCs. CD29, CD44, CD73, CD90, and CD105 were present in hTSCs. Osteogenic, adipogenic, and chondrogenic differentiation was confirmed by cytochemical staining and lineage-specific transcripts. Our results showed that specific Dulbecco modified Eagle medium F12 culture medium results in the presence of hTPC markers. hTPCs displayed lipid droplets, appropriate gene expression, and secreted dehydroepiandrosterone and estradiol. hTSCs have the ability to differentiate into mesenchymal lineages and hTPCs. This study may provide a novel in vitro model for further investigation of theca cell maturation and differentiation.

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Establishment of an in vitro culture model of theca cells from hierarchical follicles in ducks

Cited by 17 publications

References 41 publications

Comparison of growth characteristics of in vitro cultured granulosa cells from geese follicles at different developmental stages

Comparison of growth characteristics of in vitro cultured granulosa cells from geese follicles at different developmental stages

Isolation and in vitro culture of ovarian stem cells in Chinese soft‐shell turtle (Pelodiscus sinensis)

Establishment and characterization of human theca stem cells and their differentiation into theca progenitor cells

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