Comparative efficiency for in vitro transfection of goat undifferentiated spermatogonia using lipofectamine reagents and electroporation
Spermatogonial stem cells (SSC), also referred to as undifferentiated spermatogonia, are the germline stem cells responsible for continuous spermatogenesis throughout a male's life. They are, therefore, an ideal target for gene editing. Previously, SSC from animal testis have been isolated and trans...
| Autores principales: | , , , , , |
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| Formato: | Journal Article |
| Lenguaje: | Inglés |
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Informa UK Limited
2022
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| Materias: | |
| Acceso en línea: | https://hdl.handle.net/10568/119631 |
| _version_ | 1855540652186533888 |
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| author | Nakami, W. Nguhiu-Mwangi, J. Kipyegon, A.N. Ogugo, Moses Muteti, Charity Kemp, Stephen J. |
| author_browse | Kemp, Stephen J. Kipyegon, A.N. Muteti, Charity Nakami, W. Nguhiu-Mwangi, J. Ogugo, Moses |
| author_facet | Nakami, W. Nguhiu-Mwangi, J. Kipyegon, A.N. Ogugo, Moses Muteti, Charity Kemp, Stephen J. |
| author_sort | Nakami, W. |
| collection | Repository of Agricultural Research Outputs (CGSpace) |
| description | Spermatogonial stem cells (SSC), also referred to as undifferentiated spermatogonia, are the germline stem cells responsible for continuous spermatogenesis throughout a male's life. They are, therefore, an ideal target for gene editing. Previously, SSC from animal testis have been isolated and transplanted to homologous recipients resulting in the successful reestablishment of donor-derived spermatogenesis.Enhanced green fluorescent protein (eGFP) gene transfection into goat SSC was evaluated using liposomal carriers and electroporation. The cells were isolated from the prepubertal Galla goats testis cultured in serum-free defined media and transfected with the eGFP gene. Green fluorescing of SSC colonies indicated transfection.The use of lipofectamineTM stem reagent and lipofectamineTM 2000 carriers resulted in more SSC colonies expressing the eGFP gene (25.25% and 22.25%, respectively). Electroporation resulted in 15% ± 0.54 eGFP expressing SSC colonies. Furthermore, cell viability was higher in lipofectamine transfection (55% ± 0.21) as compared to electroporation (38% ± 0.14).These results indicated that lipofectamine was more effective in eGFP gene transfer into SSC. The successful transient transfection points to a possibility of transfecting transgenes into male germ cells in genetic engineering programs. |
| format | Journal Article |
| id | CGSpace119631 |
| institution | CGIAR Consortium |
| language | Inglés |
| publishDate | 2022 |
| publishDateRange | 2022 |
| publishDateSort | 2022 |
| publisher | Informa UK Limited |
| publisherStr | Informa UK Limited |
| record_format | dspace |
| spelling | CGSpace1196312024-10-03T07:41:01Z Comparative efficiency for in vitro transfection of goat undifferentiated spermatogonia using lipofectamine reagents and electroporation Nakami, W. Nguhiu-Mwangi, J. Kipyegon, A.N. Ogugo, Moses Muteti, Charity Kemp, Stephen J. animal breeding breeds genetics goats small ruminants Spermatogonial stem cells (SSC), also referred to as undifferentiated spermatogonia, are the germline stem cells responsible for continuous spermatogenesis throughout a male's life. They are, therefore, an ideal target for gene editing. Previously, SSC from animal testis have been isolated and transplanted to homologous recipients resulting in the successful reestablishment of donor-derived spermatogenesis.Enhanced green fluorescent protein (eGFP) gene transfection into goat SSC was evaluated using liposomal carriers and electroporation. The cells were isolated from the prepubertal Galla goats testis cultured in serum-free defined media and transfected with the eGFP gene. Green fluorescing of SSC colonies indicated transfection.The use of lipofectamineTM stem reagent and lipofectamineTM 2000 carriers resulted in more SSC colonies expressing the eGFP gene (25.25% and 22.25%, respectively). Electroporation resulted in 15% ± 0.54 eGFP expressing SSC colonies. Furthermore, cell viability was higher in lipofectamine transfection (55% ± 0.21) as compared to electroporation (38% ± 0.14).These results indicated that lipofectamine was more effective in eGFP gene transfer into SSC. The successful transient transfection points to a possibility of transfecting transgenes into male germ cells in genetic engineering programs. 2022-05-10 2022-05-23T10:22:08Z 2022-05-23T10:22:08Z Journal Article https://hdl.handle.net/10568/119631 en Open Access Informa UK Limited Nakami, W.N., Nguhiu-Mwangi, J., Kipyegon, A.N., Ogugo, M., Muteti, C. and Kemp, S. 2022. Comparative efficiency for in vitro transfection of goat undifferentiated spermatogonia using lipofectamine reagents and electroporation. Stem Cells and Cloning: Advances and Applications 15:11-20. |
| spellingShingle | animal breeding breeds genetics goats small ruminants Nakami, W. Nguhiu-Mwangi, J. Kipyegon, A.N. Ogugo, Moses Muteti, Charity Kemp, Stephen J. Comparative efficiency for in vitro transfection of goat undifferentiated spermatogonia using lipofectamine reagents and electroporation |
| title | Comparative efficiency for in vitro transfection of goat undifferentiated spermatogonia using lipofectamine reagents and electroporation |
| title_full | Comparative efficiency for in vitro transfection of goat undifferentiated spermatogonia using lipofectamine reagents and electroporation |
| title_fullStr | Comparative efficiency for in vitro transfection of goat undifferentiated spermatogonia using lipofectamine reagents and electroporation |
| title_full_unstemmed | Comparative efficiency for in vitro transfection of goat undifferentiated spermatogonia using lipofectamine reagents and electroporation |
| title_short | Comparative efficiency for in vitro transfection of goat undifferentiated spermatogonia using lipofectamine reagents and electroporation |
| title_sort | comparative efficiency for in vitro transfection of goat undifferentiated spermatogonia using lipofectamine reagents and electroporation |
| topic | animal breeding breeds genetics goats small ruminants |
| url | https://hdl.handle.net/10568/119631 |
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