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Wilkister Nabulindo Nakami,1â 3 James Nguhiu-Mwangi,2 Ambrose Ngâeno Kipyegon,2 Moses Ogugo,1,3 Charity Muteti,1,3 Stephen Kemp1,3 1Livestock Genetics, International Livestock Research Institute, ILRI, Nairobi, Kenya; 2Department of Clinical Studies, Faculty of Veterinary Medicine, University of Nairobi, Nairobi, Kenya; 3Centre for Tropical Livestock Genetics and Health (CTLGH), ILRI, Nairobi, KenyaCorrespondence: Wilkister Nabulindo Nakami, Livestock Genetics, International Livestock Research Institute, ILRI, 30709-00100, Nairobi, Kenya, Tel +254 711 761 459, Email wilkisterkelly@gmail.com; W.Nabulindo@cgiar.orgIntroduction: Spermatogonial stem cells (SSC), also referred to as undifferentiated spermatogonia, are the germline stem cells responsible for continuous spermatogenesis throughout a maleâs 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.Methods: 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.Results: 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).Conclusion: 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.Keywords: eGFP, culture, pre-pubertal |
