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| Content Provider | Springer Nature Link |
|---|---|
| Author | Kim, Nam Hyung Kim, Dong Hoon Kim, Yong Hee Hwang, Seongsoo Kim, Min Kyu Chung, Hak Jae Ryu, Buom Yong Kim, Bang Jin Kim, Ki Jung Jung, Sang Eun Kim, Myung Jick Kim, Byung Gak Kim, In Cheul Lee, Yong An Kim, Chul Geun Choi, Sun Ho |
| Copyright Year | 2014 |
| Abstract | Spermatogonial stem cells (SSCs) are adult stem cells that transmit genetic information from the parent to the next generation (progeny) in males, and thus, SSCs have used in germline-modification for generating transgenic animals. In this study, we demonstrated the feasibility of transgenic sperm production by employing an effective busulfan treatment method to prepare recipient pigs for the transplantation of genetically modified donor porcine SSCs. We purified SSCs from pig testis cells by sequentially employing Laminin-coated dishes and culture dishes. The purified cells were transduced with lentivirus expressing enhanced green fluorescent protein (eGFP) at a multiplicity of infection (MOI) of 6 for 9 h. eGFP transduced pig SSCs were then transplanted into the seminiferous tubules of 12 to 16-week-old recipients born to busulfan-treated sows. We obtained six recipient pigs after transplantation and maintained them for more than 6 months. The collected viable spermatozoa from 2 out of 6 recipients were positive for eGFP gene expression in polymerase chain reaction. eGFP-expressing spermatozoa appeared morphologically normal under the microscope. When spermatozoa from these recipients were used for intra cytoplasmic sperm injection, eGFP expression could be detected in the embryos. Furthermore, eGFP colonies were derived from donor-transduced SSCs observed in the recipients’ testes. In summary, we demonstrated the successful production of functional-transgenic spermatozoa by transplantation of porcine SSCs where the transgenic was transduced by employing the lentiviral vector system. |
| Starting Page | 458 |
| Ending Page | 466 |
| Page Count | 9 |
| File Format | |
| ISSN | 17382696 |
| Journal | Tissue Engineering and Regenerative Medicine |
| Volume Number | 11 |
| Issue Number | 6 |
| e-ISSN | 22125469 |
| Language | English |
| Publisher | Korean Tissue Engineering and Regenerative Medicine Society |
| Publisher Date | 2014-11-03 |
| Publisher Place | Dordrecht |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Biomedical Engineering spermatogonial stem cell lentiviral transduction transplantation transgenic sperm Biomedicine general pig Cell Biology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Medicine Biomedical Engineering |
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