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Establishment of an electroporation-mediated gene delivery system in porcine spermatogonial stem cells

  • Min Seong Kim
  • Min Hee Park
  • Ji Eun Park
  • Jung Im Yun
  • Jung Hoon Choi
  • Eunsong Lee
  • Seung Tae LeeEmail author
Article
  • 17 Downloads

Abstract

Spermatogonial stem cells (SSCs) are a useful tool for the generation of genetically modified transgenic sperm. As a result, the transfer of specific genes into the cytoplasm of SSCs is crucial for the successful generation of transgenic sperm. Here, we report electroporation conditions optimized for SSCs derived from the porcine testis. The highest transfection efficiency and cell viability were observed in porcine SSCs transfected with 1 μg transgenic vector with a single electric pulse from an electroporator at a voltage of 100 V and a capacitor setting of 250 μF. The transfection efficiency and cell viability were constant regardless of the size of the transgenic vector. Furthermore, we did not detect loss of spermatozoa differentiation potential in the transfected porcine SSCs. From these results, we confirm that this electroporation-based gene delivery system can effectively introduce foreign DNA into the genome of porcine SSCs without any loss of the original porcine SSC characteristics, which will be important in the generation of mosaicism-free transgenic pigs produced from transgenic porcine sperm.

Keywords

Pig Spermatogonial stem cells Transfection Gene delivery system Electroporation 

Notes

Acknowledgements

We thank Joohyeong Lee and Keun Cheon Kim for assistance of IVF and cryostat sectioning, respectively.

Funding information

This research was supported by the Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries (IPET) through Agri-Bioindustry Technology Development Program, funded by Ministry of Agriculture, Food and Rural Affairs (MAFRA) (IPET117042-3), Republic of Korea.

Compliance with ethical standards

All of the animal experimental procedures were approved by the Institutional Animal Care and Use Committee of Kangwon National University (Approval No. KW-131106-1) and were performed in accordance with the Animal Care and Use Guidelines of Kangwon National University (Kangwon-do, South Korea).

Conflict of interest

The authors declare that they have no conflict of interests.

Supplementary material

11626_2019_326_MOESM1_ESM.docx (48 kb)
ESM 1 (DOCX 47 kb)

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Copyright information

© The Society for In Vitro Biology 2019

Authors and Affiliations

  • Min Seong Kim
    • 1
  • Min Hee Park
    • 1
  • Ji Eun Park
    • 1
  • Jung Im Yun
    • 2
  • Jung Hoon Choi
    • 3
  • Eunsong Lee
    • 3
  • Seung Tae Lee
    • 1
    • 4
    Email author
  1. 1.Department of Animal Life ScienceKangwon National UniversityChuncheonSouth Korea
  2. 2.Division of Animal Resource ScienceKangwon National UniversityChuncheonSouth Korea
  3. 3.College of Veterinary MedicineKangwon National UniversityChuncheonSouth Korea
  4. 4.Department of Applied Animal Science, College of Animal Life ScienceKangwon National UniversityChuncheonSouth Korea

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