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

OBJECTIVES: In vitro differentiation of oocytes from female germline stem cells (FGSCs) has exciting potential applications for reproductive medicine. Some researchers have attempted to reveal the in vitro differentiation capacity of FGSCs. However, no systematic comparative study of in vitro differ...

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Autores principales: Zou, Kang, Wang, Jian, Bi, Haiwei, Zhang, Yabin, Tian, Xueli, Tian, Ning, Ma, Wanyun, Wu, Ji
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6430485/
https://www.ncbi.nlm.nih.gov/pubmed/30334302
http://dx.doi.org/10.1111/cpr.12530
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author Zou, Kang
Wang, Jian
Bi, Haiwei
Zhang, Yabin
Tian, Xueli
Tian, Ning
Ma, Wanyun
Wu, Ji
author_facet Zou, Kang
Wang, Jian
Bi, Haiwei
Zhang, Yabin
Tian, Xueli
Tian, Ning
Ma, Wanyun
Wu, Ji
author_sort Zou, Kang
collection PubMed
description OBJECTIVES: In vitro differentiation of oocytes from female germline stem cells (FGSCs) has exciting potential applications for reproductive medicine. Some researchers have attempted to reveal the in vitro differentiation capacity of FGSCs. However, no systematic comparative study of in vitro differentiation conditions has been performed for murine FGSCs (mFGSCs). MATERIALS AND METHODS: mFGSCs line was cultured under five different conditions for in vitro differentiation. RT‐PCR was performed to detect the expression of Oct4, Fragilis, Blimp1, Mvh, Scp3 and Zp3. Immunofluorescence was carried out to test the expression of Mvh, Fragilis and Zp3. Two‐photon laser‐scanning microscope was used to analyze nucleus‐plasma ratio, and the proportion of chromatin of GV oocytes differentiated from mFGSCs in vitro (IVD‐GVO), GV oocytes from in vivo (GVO) and mFGSCs. RESULTS: RT‐PCR and immunofluorescence showed that mFGSC line expressed germ cell‐specific markers, but not a meiosis‐specific marker. By evaluating five different in vitro differentiation conditions, condition 5, which included a hanging drop procedure and co‐culture of mFGSCs with granulosa cells, was shown to be optimal. mFGSCs could be successfully differentiated into germinal vesicle (GV) ‐stage oocytes under this condition. 3D observation revealed that both the nucleus‐plasma ratio and proportion of chromatin were not significantly different between IVD‐GVO and GVO. CONCLUSION: We evaluated five in vitro differentiation conditions for mFGSCs and successfully differentiate mFGSCs into GV‐stage oocytes using a three‐step differentiation process.
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spelling pubmed-64304852020-03-13 Comparison of different in vitro differentiation conditions for murine female germline stem cells Zou, Kang Wang, Jian Bi, Haiwei Zhang, Yabin Tian, Xueli Tian, Ning Ma, Wanyun Wu, Ji Cell Prolif Original Articles OBJECTIVES: In vitro differentiation of oocytes from female germline stem cells (FGSCs) has exciting potential applications for reproductive medicine. Some researchers have attempted to reveal the in vitro differentiation capacity of FGSCs. However, no systematic comparative study of in vitro differentiation conditions has been performed for murine FGSCs (mFGSCs). MATERIALS AND METHODS: mFGSCs line was cultured under five different conditions for in vitro differentiation. RT‐PCR was performed to detect the expression of Oct4, Fragilis, Blimp1, Mvh, Scp3 and Zp3. Immunofluorescence was carried out to test the expression of Mvh, Fragilis and Zp3. Two‐photon laser‐scanning microscope was used to analyze nucleus‐plasma ratio, and the proportion of chromatin of GV oocytes differentiated from mFGSCs in vitro (IVD‐GVO), GV oocytes from in vivo (GVO) and mFGSCs. RESULTS: RT‐PCR and immunofluorescence showed that mFGSC line expressed germ cell‐specific markers, but not a meiosis‐specific marker. By evaluating five different in vitro differentiation conditions, condition 5, which included a hanging drop procedure and co‐culture of mFGSCs with granulosa cells, was shown to be optimal. mFGSCs could be successfully differentiated into germinal vesicle (GV) ‐stage oocytes under this condition. 3D observation revealed that both the nucleus‐plasma ratio and proportion of chromatin were not significantly different between IVD‐GVO and GVO. CONCLUSION: We evaluated five in vitro differentiation conditions for mFGSCs and successfully differentiate mFGSCs into GV‐stage oocytes using a three‐step differentiation process. John Wiley and Sons Inc. 2018-10-17 /pmc/articles/PMC6430485/ /pubmed/30334302 http://dx.doi.org/10.1111/cpr.12530 Text en © 2018 The Authors Cell Proliferation Published by John Wiley & Sons Ltd This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Articles
Zou, Kang
Wang, Jian
Bi, Haiwei
Zhang, Yabin
Tian, Xueli
Tian, Ning
Ma, Wanyun
Wu, Ji
Comparison of different in vitro differentiation conditions for murine female germline stem cells
title Comparison of different in vitro differentiation conditions for murine female germline stem cells
title_full Comparison of different in vitro differentiation conditions for murine female germline stem cells
title_fullStr Comparison of different in vitro differentiation conditions for murine female germline stem cells
title_full_unstemmed Comparison of different in vitro differentiation conditions for murine female germline stem cells
title_short Comparison of different in vitro differentiation conditions for murine female germline stem cells
title_sort comparison of different in vitro differentiation conditions for murine female germline stem cells
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6430485/
https://www.ncbi.nlm.nih.gov/pubmed/30334302
http://dx.doi.org/10.1111/cpr.12530
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