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

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 fo...

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Published inIn vitro cellular & developmental biology. Animal Vol. 55; no. 3; pp. 177 - 188
Main Authors Kim, Min Seong, Park, Min Hee, Park, Ji Eun, Yun, Jung Im, Choi, Jung Hoon, Lee, Eunsong, Lee, Seung Tae
Format Journal Article
LanguageEnglish
Published New York Springer Science & Business Media LLC 01.03.2019
Springer US
Society for In Vitro Biology
Subjects
Animal Genetics and Genomics
Biomedical and Life Sciences
BIOTECHNOLOGY
Cell Biology
Cell Culture
Cell viability
Cytoplasm
Deoxyribonucleic acid
Developmental Biology
DNA
Electroporation
Gene transfer
Genetic modification
Genomes
Life Sciences
Mosaicism
Sperm
Spermatozoa
Stem cell transplantation
Stem Cells
Swine
Transfection
Electroporation
Transfection
Gene delivery system
Spermatogonial stem cells
Pig
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Summary: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.
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ISSN:1071-2690
1543-706X
DOI:10.1007/s11626-019-00326-7