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In vitro propagation of male germline stem cells from piglets

  • Stem Cell Biology
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

To study the effects of serum and growth factors on propagation of porcine male germline stem cells (MGSCs) in vitro and develop a culture system for these stem cells.

Methods

Fresh testicular cells from neonatal piglets were obtained by mechanical dissociation and collagenase-trypsin digestion. After differential plating, non-adhering cells were cultured in media supplemented with different concentrations of serum (0, 1 %, 2 %, 5 %, 10 %). After 10 days of primary culture, the cells were maintained in media supplemented with different concentrations of growth factors (basic fibroblast growth factor and epidermal growth factor at 1, 5, 10 ng/ml). The number of MGSC-derived colonies with different sizes was determined in each treatment to assess the effects of serum concentrations and growth factors.

Results

The number of MGSC-derived colonies was significantly higher in the presence of 1 % rather than 10 % fetal bovine serum (FBS). Basic fibroblast growth factor (bFGF) at 1, 5 ng/ml and epidermal growth factor (EGF) at 5, 10 ng/ml significantly promoted colony formation. Immunocytochemistry, reverse transcriptase-polymerase chain reaction (RT-PCR) and xenotransplantation assays demonstrated the presence of functional stem cells in cultured cell population.

Conclusions

In vitro propagation of porcine MGSCs could be maintained in the presence of 1 % FBS and supplementation of growth factors for 1 month.

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Acknowledgments

We thank Yangling Guangming pig farm for providing porcine testis tissues. This work was supported by the National Natural Science Foundation of China (Grant No. 31072029, No. 31272439), and the Scientific Research Foundation for the Returned Overseas Chinese Scholars from Northwest A&F University.

Declaration of interest

The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported.

Author information

Authors and Affiliations

  1. College of Animal Science and Technology, Northwest A&F University, 22 Xi-nong Road, Yangling, Shaanxi, 712100, People’s Republic of China

    Yi Zheng, Xiue Tian, Yaqing Zhang, Jinzhou Qin, Junhui An & Wenxian Zeng

  2. Shaanxi Center of Stem Cells Engineering and Technology, Yangling, Shaanxi, 712100, China

    Yi Zheng, Xiue Tian, Yaqing Zhang, Jinzhou Qin, Junhui An & Wenxian Zeng

  3. Key Lab for Biotechnology of Agriculture Ministry of China, Yangling, Shaanxi, 712100, China

    Yi Zheng, Xiue Tian, Yaqing Zhang, Jinzhou Qin, Junhui An & Wenxian Zeng

Authors
  1. Yi Zheng
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  2. Xiue Tian
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  3. Yaqing Zhang
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  4. Jinzhou Qin
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  5. Junhui An
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  6. Wenxian Zeng
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Corresponding author

Correspondence to Wenxian Zeng.

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Capsule

This study showed the optimum concentrations of serum and growth factors for propagation of porcine male germline stem cells (MGSCs) in vitro and finally established a culture system for these stem cells.

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Zheng, Y., Tian, X., Zhang, Y. et al. In vitro propagation of male germline stem cells from piglets. J Assist Reprod Genet 30, 945–952 (2013). https://doi.org/10.1007/s10815-013-0031-0

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  • DOI: https://doi.org/10.1007/s10815-013-0031-0

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