Improvement in Isolation and Identification of Mouse Oogonial Stem Cells

Female germline stem cells (FGSCs) or oogonial stem cells (OSCs) have the capacity to generate newborn oocytes and thus open a new door to fight ovarian aging and female infertility. However, the production and identification of OSCs are difficult for investigators. Rare amount of these cells in the...

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Main Authors: Zhiyong Lu, Meng Wu, Jinjin Zhang, Jiaqiang Xiong, Jing Cheng, Wei Shen, Aiyue Luo, Li Fang, Shixuan Wang
Format: Article
Language:English
Published: Hindawi Limited 2016-01-01
Series:Stem Cells International
Online Access:http://dx.doi.org/10.1155/2016/2749461
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spelling doaj-538630a2e82c4e009d6e98ef3dcc8ad92020-11-24T23:10:32ZengHindawi LimitedStem Cells International1687-966X1687-96782016-01-01201610.1155/2016/27494612749461Improvement in Isolation and Identification of Mouse Oogonial Stem CellsZhiyong Lu0Meng Wu1Jinjin Zhang2Jiaqiang Xiong3Jing Cheng4Wei Shen5Aiyue Luo6Li Fang7Shixuan Wang8Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Road, Wuhan, Hubei 430030, ChinaDepartment of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Road, Wuhan, Hubei 430030, ChinaDepartment of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Road, Wuhan, Hubei 430030, ChinaDepartment of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Road, Wuhan, Hubei 430030, ChinaDepartment of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Road, Wuhan, Hubei 430030, ChinaDepartment of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Road, Wuhan, Hubei 430030, ChinaDepartment of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Road, Wuhan, Hubei 430030, ChinaDepartment of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Road, Wuhan, Hubei 430030, ChinaDepartment of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Road, Wuhan, Hubei 430030, ChinaFemale germline stem cells (FGSCs) or oogonial stem cells (OSCs) have the capacity to generate newborn oocytes and thus open a new door to fight ovarian aging and female infertility. However, the production and identification of OSCs are difficult for investigators. Rare amount of these cells in the ovary results in the failure of the acquisition of OSCs. Furthermore, the oocyte formation by OSCs in vivo was usually confirmed using tissue sections by immunofluorescence or immunohistochemistry in previous studies. STO or MEF feeder cells are derived from mouse, not human. In our study, we modified the protocol. The cells were digested from ovaries and cultured for 2-3 days and then were purified by magnetic-activated cell sorting (MACS). The ovaries and fetus of mice injected with EGFP-positive OSCs were prepared and put on the slides to directly visualize oocyte and progeny formation under microscope. Additionally, the human umbilical cord mesenchymal stem cells (hUC-MSCs) were also used as feeder cells to support the proliferation of OSCs. The results showed that all the modified procedures can significantly improve and facilitate the generation and characterization of OSCs, and hUC-MSCs as feeder will be useful for isolation and proliferation of human OSCs avoiding contamination from mouse.http://dx.doi.org/10.1155/2016/2749461
collection DOAJ
language English
format Article
sources DOAJ
author Zhiyong Lu
Meng Wu
Jinjin Zhang
Jiaqiang Xiong
Jing Cheng
Wei Shen
Aiyue Luo
Li Fang
Shixuan Wang
spellingShingle Zhiyong Lu
Meng Wu
Jinjin Zhang
Jiaqiang Xiong
Jing Cheng
Wei Shen
Aiyue Luo
Li Fang
Shixuan Wang
Improvement in Isolation and Identification of Mouse Oogonial Stem Cells
Stem Cells International
author_facet Zhiyong Lu
Meng Wu
Jinjin Zhang
Jiaqiang Xiong
Jing Cheng
Wei Shen
Aiyue Luo
Li Fang
Shixuan Wang
author_sort Zhiyong Lu
title Improvement in Isolation and Identification of Mouse Oogonial Stem Cells
title_short Improvement in Isolation and Identification of Mouse Oogonial Stem Cells
title_full Improvement in Isolation and Identification of Mouse Oogonial Stem Cells
title_fullStr Improvement in Isolation and Identification of Mouse Oogonial Stem Cells
title_full_unstemmed Improvement in Isolation and Identification of Mouse Oogonial Stem Cells
title_sort improvement in isolation and identification of mouse oogonial stem cells
publisher Hindawi Limited
series Stem Cells International
issn 1687-966X
1687-9678
publishDate 2016-01-01
description Female germline stem cells (FGSCs) or oogonial stem cells (OSCs) have the capacity to generate newborn oocytes and thus open a new door to fight ovarian aging and female infertility. However, the production and identification of OSCs are difficult for investigators. Rare amount of these cells in the ovary results in the failure of the acquisition of OSCs. Furthermore, the oocyte formation by OSCs in vivo was usually confirmed using tissue sections by immunofluorescence or immunohistochemistry in previous studies. STO or MEF feeder cells are derived from mouse, not human. In our study, we modified the protocol. The cells were digested from ovaries and cultured for 2-3 days and then were purified by magnetic-activated cell sorting (MACS). The ovaries and fetus of mice injected with EGFP-positive OSCs were prepared and put on the slides to directly visualize oocyte and progeny formation under microscope. Additionally, the human umbilical cord mesenchymal stem cells (hUC-MSCs) were also used as feeder cells to support the proliferation of OSCs. The results showed that all the modified procedures can significantly improve and facilitate the generation and characterization of OSCs, and hUC-MSCs as feeder will be useful for isolation and proliferation of human OSCs avoiding contamination from mouse.
url http://dx.doi.org/10.1155/2016/2749461
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