Chinese Journal of Tissue Engineering Research ›› 2014, Vol. 18 ›› Issue (19): 3088-3093.doi: 10.3969/j.issn.2095-4344.2014.19.022

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Establishment and characterization of CD-1 neonatal female germline stem cell lines

Bi Hai-wei1, Wu Ji1, 2   

  1. 1Key Laboratory for the Genetics of Developmental & Neuropsychiatric Disorders (Ministry of Education), Bio-X Institutes, Shanghai Jiao Tong University, Shanghai 200240, China
    2Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • Revised:2014-03-27 Online:2014-05-07 Published:2014-05-07
  • Contact: Wu Ji, Professor, Doctoral supervisor, Key Laboratory for the Genetics of Developmental & Neuropsychiatric Disorders (Ministry of Education), Bio-X Institutes, Shanghai Jiao Tong University, Shanghai 200240, China; Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • About author:Bi Hai-wei, Master, Key Laboratory for the Genetics of Developmental & Neuropsychiatric Disorders (Ministry of Education), Bio-X Institutes, Shanghai Jiao Tong University, Shanghai 200240, China
  • Supported by:

    the National Basic Research Program of China, No. 2013CB967401; the National Natural Science Foundation of China, No. 81370675

Abstract:

BACKGROUND: In traditional view, the germ cells in mammal ovaries were considered to lose the capacity of self-renew after birth. However, recent studies have showed the existence of female germline stem cells (FGSCs) which take the responsibility for regenerating oocytes in the ovaries of postnatal mammals.
OBJECTIVE: To establish the FGSC line and to explore the biological characterization of the FGSC line from neonatal CD-1 mouse ovaries.
METHODS: Using two-step enzymatic digestion and immunomagnetic purification, Fragilis-positive cells were isolated from neonatal CD-1 mice ovaries and subjected to long-term culture. Then their biological characteristics were studies with RT-PCR, fluorescent immunocytochemistry and cytogenetic analysis.
RESULTS AND CONCLUSION: FGSCs from neonatal CD-1 mice aged 3-5 days were isolated and long-term cultured for 70 passages to establish a cell line. This cell line expressed Mvh, Dazl, Oct-4, Stella, Blimp1, Fragilis, and was proven to be mitotically active germ cells by BrdU/Fragilis dual immunofluorescence analysis. Moreover, cytogenetic analysis by Gimsa staining confirmed their normal karyotype. In all, these results indicate the existence of FGSCs with mitotic activity in neonatal CD-1 mouse ovaries.


中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程


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Key words: stem cells, reproduction, germ cells, cell proliferation, oocytes, biology

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