Abstract
A cell population enriched with type A spermatogonia has been isolated from the boar testes. Cell types occurring during isolation were morphologically characterized, factors maintaining the cultured spermatogonia in the undifferentiated state were studied, and these cells were transferred to sterile recipients preliminarily treated with busulfan. The cells of spermatogenic epithelium cultivated in vitro for 24 h were used for transfer experiments. The transfer efficiency was estimated within 27 and 29 days according to the histological picture of the testes and the isolated cultures. Spermatogenic cells at various developmental stages and a few Sertoli cells and spermatozoa were found on sections and in cell suspensions. Sperm samples could be taken from recipient boars within nine months after the transfer. Microsatellite analysis of DNA showed the endogenous pattern of spermatogenesis. Thus, it was shown that spermatogenic donor cells can restore and maintain spermatogenesis of a recipient for at least 30 days. However, the donor cells were fully forced by the recipient reserve cells, type A0 spermatogonia, within eight to nine months.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Brinster, R. and Avarbock, M., Germline Transmission of Donor Haplotype Following Spermatogonial Transplantation, Proc. Natl. Acad. Sci. USA, 1994, vol. 91, pp. 11303–11307.
Brinster, R. and Zimmermann, J., Spermatogenesis Following Male Germ-Cell Transplantation, Proc. Natl. Acad. USA, 1994, vol. 91, pp. 11298–11302.
Bucci, L.R. and Meistrich, M.L., Effects of Busulfan on Murine Spermatogenesis: Cytotoxicity, Sterility, Sperm Abnormalities, and Dominant Lethal Mutation, Mutat. Res., 1987, vol. 176, pp. 259–268.
Dirami, G., Ravindranath, N., Pursel, V., et al., Effects of Stem Cell Factor and Granulocyte Macrophage-Colony Stimulating Factor on Survival of Porcine Type A Spermatogonia Cultured in KSOM, Biol. Reprod., 1999, vol. 61, pp. 225–230.
Dobrinski, I., Avarbock, M.R., and Brinster, R.L., Transplantation of Germ Cells from Rabbits and Dogs Into Mouse Testes, Biol. Reprod., 1999, vol. 61, pp. 1331–1339.
Franca, L., Ogawa, T., Avarbock, M.R., et al., Germ Cell Genotype Controls Cell Cycle During Spermatogenesis in the Rat, Biol. Reprod., 1998, vol. 59, pp. 1371–1377.
Honaramooz, A., Megee, S.O., and Dobrinski, I., Germ Cell Transplantation in Pigs, Biol. Reprod., 2002, vol. 66, pp. 21–28.
Izadyar, F., Spierenberg, G.T., Creemers, L.B., et al., Isolation and Purification of Type A Spermatogonia from the Bovine Testis, Reproduction, 2002, vol. 124, pp. 85–94.
Jiang, F.X., Behaviour of Spermatogonia Following Recovery from Busulfan Treatment in the Rat, Anat. Embryol., 1998, vol. 198, pp. 53–61.
Kanatsu-Shinohara, M., Ogura, A., Ikegawa, M., et al., Adenovirus-Mediated Gene Delivery and in vitro Microinsemination Produce Offspring from Infertile Male Mice, Proc. Natl. Acad. Sci. USA, 2002, vol. 99, pp. 1383–1388.
Kim, J., Jung-Ha, H., Lee, H., et al., Development of a Positive Method for Male Stem Cell-Mediated Gene Transfer in Mouse and Pig, Mol. Reprod. Devel., 1997, vol. 46, pp. 515–526.
Kostereva, N.V., Korjikova, S.V., Savchenkova, I.P., et al., Dynamics of Spermatogenesis in Pigs as Seen in Samples of Fixed Tissues, Mater. nauch.-praktich. konf. po probleme “Povyshenie konkurentosposobnosti zhivotnovodstva i zadachi kadrovogo obespecheniya” (Materrials of Scientific-Practical Conference “Improvement of Competitive Capacity of Animal Husbandry and Problems of Staff”). Bykovo: Izd-e RA, MZh, 2001, pp. 166–168.
Kubota, H., Avarbock, M.R., and Brinster, L., Spermatogonial Stem Cells Share Some, but not All, Phenotypic and Functional Characteristics with Other Stem Cells, Proc. Natl. Acad. Sci. USA, 2003, vol. 100, pp. 6487–6492.
Meachem, S., von Schönfeldt, V., and Schlatt, S., Spermatogonia: Stem Cells with a Great Perspective, Reproduction, 2001, vol. 121, pp. 825–834.
Nagano, M., Avarbock, M., Lenida, E., et al., Culture of Mouse Spermatogonial Stem Cells, Tiss. Cell, 1998, vol. 30, pp. 389–397.
Nagano, M., Brinster, C.J., Orwig, K.E., et al., Transgenic Mice Produced by Retroviral Transduction of Male Germ-Line Stem Cells, Proc. Natl. Acad. Sci. USA, 2001a, vol. 98, pp. 13090–13095.
Nagano, M., McCarrey, J.R., and Brinster, R.L., Primate Spermatogonial Stem Cell Colonize Mouse Testes, Biol. Reprod., 2001b, vol. 64, pp. 1409–1416.
Ogawa, T., Arechaga, J.M., Avarbock, M.R., et al., Transplantation of Testis Germinal Cells Into Mouse Seminiferous Tubules, Int. J. Devel. Biol., 1997, vol. 41, pp. 111–122.
Ogawa, T., Dobrinski, I., Avarbock, M.R., et al., Leuorolide, a Gonadotropin — Releasing Hormone Agonist, Enhances Colonization after Spermatogonial Transplantation Into Mouse Testes, Tiss. Cell, 1998, vol. 30, pp. 583–588.
Ogawa, T., Dobrinski, I., Avarbock, M.R., et al., Xenogenic Spermatogenesis Following Transplantation of Hamster Germ Cells to Mouse Testes, Biol. Reprod., 1999, vol. 60, pp. 515–521.
Orwig, K.E., Ryu, B.Y., Avarbock, M.R., et al., Male Germ-Line Stem Cell Potential Is Predicted by Morphology of Cells in Neonatal Rat Testes, Proc. Natl. Acad. Sci. USA, 2002, vol. 99, pp. 11706–11711.
Parriera, G., Ogawa, T., Avarbock, M.R., et al., Development of Germ Cell Transplants in Mice, Biol. Reprod., 1998, vol. 59, pp. 1360–1370.
Romeis, B., Mikroskopicheskaya tekhnika (Microscopic Technique), Moscow: Inostrannaya Literatura, 1953.
de Rooij, D.G., Proliferation and Differentiation of Spermatogonial Stem Cells, Reproduction, 2001, vol. 121, pp. 347–354.
Roskin, G.I., Mikroskopicheskaya tekhnika (Microscopic Technique), Moscow: Sov. nauka, 1946.
Savchenkova, I.P., Embrional’nye stvolovye kletki v biologii: nastoyashchee i budushchee (Embryonic Stem Cells in Biology: Present and Future), Dubrovitsy, 1999.
Savchenkova, I.P., Embryonic Stem Cells in Biology and Biotechnology, Zhivotnaya kletka v kul’ture (Animal Cell in Culture), Moscow: Komp. “Sputnik,” 2000.
Savchenkova, I.P., Pridantseva, T.A., Sergeev, H.V., et al., Isolation and Purification of Primordial Germ Cells of Mouse Embryos, S.-Kh. Biol., 2000, no. 2, pp. 119–121.
Shinohara, T., Orwig, K.E., Avarbock, M.R., et al., Spermatogonial Stem Cell Enrichment by Multiparameter Selection of Mouse Testis Cells, Proc. Natl. Acad. Sci. USA, 2000, vol. 97, pp. 8346–8351.
Tegelenbosch, R.A. and de Rooij, D.G., A Quantitative Study of Spermatogonial Multiplication and Stem Cell Renewal in the C3H/101 F1 Hybrid Mouse, Mutat. Res., 1993, vol. 290, pp. 193–200.
Vecino, P., Uranga, J.A., and Arechaga, J., Suppression of Spermatogenesis for Cell Transplantation in Adult Mice, Protoplasma, 2001, vol. 217, pp. 191–198.
Yamazaki, Y., Fujimoto, H., Ando, H., et al., in vivo Gene Transfer to Mouse Spermatogenic Cells by Deoxyribonucleic Acid Injection Into Seminiferous Tubules and Subsequent Electroporation, Biol. Reprod., 1998, vol. 59, pp. 1439–1444.
Author information
Authors and Affiliations
Additional information
Original Russian Text © I.P. Savchenkova, S.V. Korjikova, N.V. Kostereva, L.K. Ernst, 2006, published in Ontogenez, 2006, Vol. 37, No. 4, pp. 292–300.
Rights and permissions
About this article
Cite this article
Savchenkova, I.P., Korjikova, S.V., Kostereva, N.V. et al. Cultivation and transfer of porcine type A spermatogonia. Russ J Dev Biol 37, 242–249 (2006). https://doi.org/10.1134/S1062360406040060
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1134/S1062360406040060