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Long-term follow-up of porcine male germ cells transplanted into mouse testes

Published online by Cambridge University Press:  01 November 2007

Y-J. Choi ,
J-K. Park ,
M-S. Lee ,
J.D. Ahn ,
K-C. Hwang ,
H. Song  and
J-H. Kim
Y-J. Choi
Affiliation:
Department of Animal Biotechnology, College of Animal Bioscience and Technology, KonKuk University, Seoul 143–701, Korea.
J-K. Park
Affiliation:
Animal Biotechnology Division, National Livestock Research Institute, RDA, Suwon 441–706, Korea.
M-S. Lee
Affiliation:
Department of Dairy Science, Division of Applied Science, Gyeongsang National University, Jin-Ju, GyeongNam 660–701, Korea.
J.D. Ahn
Affiliation:
CHO-A Biotechnology Research Institute, CHO-A Pharm. Co. Ltd, Kwangjin-Gu, Seoul 140–701, Korea.
K-C. Hwang
Affiliation:
Department of Dairy Science, Division of Applied Science, Gyeongsang National University, Jin-Ju, GyeongNam 660–701, Korea.
H. Song
Affiliation:
Department of Animal Science, College of Natural Science, KonKuk University, Chung-ju, Chung-buk 380–701, Korea.
J-H. Kim*
Affiliation:
Department of Animal Biotechnology, College of Animal Bioscience and Technology, KonKuk University, Seoul 143–701, Korea.
*
All correspondence to: Jin-Hoi Kim, Department of Animal Biotechnology, College of Animal Bioscience and Technology, Kon-Kuk University, Seoul 143–701, Korea. Tel: +82 2 450 3687. Fax: +82 2 2201 3353. e-mail: jhkim541@konkuk.ac.kr
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Summary

This study investigated the effect of increased phylogenetic distance on the outcome of spermatogonial transplantation, with porcine donors and mice recipients. It was designed to develop a technique for detecting foreign donor cells in recipient animals. Porcine male germ cells were harvested from postnatal male testes and incubated with the lipophilic membrane dye PKH-26. For transplantation, approximately 106 PKH-26-labelled porcine male germ cells were injected into the efferent ducts of mouse testes. Animals were sacrificed at post-graft days 1, 10, 30, 45, 60 and 150 (n = 5 each). Serial frozen sections of explanted testes were prepared for detecting labelled cells. Transplanted porcine donor cells were easily detected in the recipient tubules for 8 weeks. After transplantation, we could detect both incorporation into the basement membrane and differentiation of grafted porcine donor cells by our double detection system, using PKH staining and slide PCR. However, our RT-PCR and apoptosis results revealed that most of the grafted porcine male donor cells could not differentiate past early-meiotic spermatocytes. We could induce partial differentiation of xenografted porcine donor cells in mouse testes, but not full induction of spermatogenesis. We have developed a very reliable technique for detecting foreign donor cells in recipient animals using a combination of PKH staining and slide PCR methods. Our results provide a valuable experimental model for applying and evaluating this technology in other species.

Keywords

PKH-26 stainSlide PCRXenotransplantation
Type
Research Article
Information
Zygote , Volume 15 , Issue 4 , November 2007 , pp. 325 - 335
Copyright
Copyright © Cambridge University Press 2007

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References

Brinster, C.J., Ryu, B.Y., Avarbock, M.R., Karagenc, L., Brinster, R.L. & Orwig, K.E. (2003). Restoration of fertility by germ cell transplantation requires effective recipient preparation. Biol. Reprod. 69, 412–20.CrossRefGoogle ScholarPubMed
Brinster, R.L. & Avarbock, M.R. (1994). Germline transmission of donor haplotype following spermatogonial transplantation. Proc. Natl. Acad. Sci. USA 91, 11303–7.CrossRefGoogle ScholarPubMed
Choi, Y.J., Ok, D.W., Kwon, D.N., Chung, J.I., Kim, H.C., Yeo, S.M., Kim, T., Seo, H.G. & Kim, J.H. (2004). Murine male germ cell apoptosis induced by busulfan treatment correlates with loss of c-kit-expression in a Fas/FasL- and p53-independent manner. FEBS Lett 575, 41–51.CrossRefGoogle Scholar
Choi, Y.J., Song, H., Kwon, D.N., Cho, S.K., Kang, S.J., Yoe, S.M., Kim, H.C., Lee, H.T., Park, C. & Kim, J.H. (2006). Significant IgG-immunoreactivity of the spermatogonia of the germ cell-depleted testis after busulfan treatment. Animal Reprod. Sci. 91, 317–35.CrossRefGoogle ScholarPubMed
Dobrinski, I., Avarbock, M.R. & Brinster, R.L. (1999). Transplantation of germ cells from rabbits and dogs into mouse testes. Biol. Reprod. 61, 1331–9.CrossRefGoogle ScholarPubMed
Dobrinski, I., Avarbock, M.R. & Brinster, R.L. (2000). Germ cell transplantation from large domestic animals into mouse testes. Mol. Reprod. Devel. 57, 270–9.3.0.CO;2-Z>CrossRefGoogle ScholarPubMed
Franca, L.R. & Cardoso, F.M. (1998). Duration of spermatogenesis and sperm transit time through the epididymis in the Piau boar. Tissue Cell 30, 573–82.CrossRefGoogle ScholarPubMed
Horan, P.K., Melnicoff, M.J., Jensen, B.D. & Slezak, S.E. (1990). Fluorescent cell labeling for in vivo and in vitro cell tracking. Methods Cell Biol. 33, 469–90.CrossRefGoogle ScholarPubMed
Horan, P.K. & Slezak, S.E. (1989). Stable cell membrane labelling. Nature 340, 167–8.CrossRefGoogle ScholarPubMed
Johnston, D.S., Russell, L.D. & Griswold, M.D. (2000). Advances in spermatogonial stem cell transplantation. Reviews Reprod. 5, 183–8.CrossRefGoogle ScholarPubMed
Kim, J.H., Jung-Ha, H.S., Lee, H.T. & Chung, K.S. (1997). Development of a positive method for male stem cell-mediated gene transfer in mouse and pig. Mol. Reprod. Devel. 46, 515–26.3.0.CO;2-V>CrossRefGoogle ScholarPubMed
Kubota, H., Avarbock, M.R. & Brinster, R.L. (2003). Spermatogonial stem cells share some, but not all, phenotypic and functional characteristics with other stem cells. Proc. Natl Acad. Sci. USA 100, 6487–92.CrossRefGoogle Scholar
Kwon, D.N., Seo, H.G. & Kim, J.H. (2002). Cloning, sequencing and expression analysis of the porcine uroplakin II gene. Biochem. Biophys. Res. Commun. 293, 862–9.CrossRefGoogle ScholarPubMed
Nagano, M., Patrizio, P. & Brinster, R.L. (2002). Long-term survival of human spermatogonial stem cells in mouse testes. Fertil. Steril. 78, 1225–1233.CrossRefGoogle ScholarPubMed
Oatley, J.M., de Avila, D.M., McLean, D.J., Griswold, M.D. & Reeves, J.J. (2002). Transplantation of bovine germinal cells into mouse testes. J. Anim. Sci. 80, 1925–31.CrossRefGoogle ScholarPubMed
Ogawa, T., Dobrinski, I. & Brinster, R.L. (1999). Recipient preparation is critical for spermatogonial transplantation in the rat. Tissue Cell 31, 461–72.CrossRefGoogle ScholarPubMed
Ohta, H. & Wakayama, T. (2004). Full-term development of offspring using round spermatids produced ectopically from fetal male germ cells. J. Reprod. Devel. 50, 429–37.CrossRefGoogle ScholarPubMed
Olive, V. & Cuzin, F. (2005). The spermatogonial stem cell: from basic knowledge to transgenic technology. Int. J. Biochem. Cell Biol. 37, 246–50.CrossRefGoogle ScholarPubMed
Orwig, K.E. & Schlatt, S. (2005). Cryopreservation and transplantation of spermatogonia and testicular tissue for preservation of male fertility. J. Natl Cancer Inst. 34, 51–6.CrossRefGoogle Scholar
Russell, L.D. & Brinster, R.L. (1996). Ultrastructural observations of spermatogenesis following transplantation of rat testis cells into mouse seminiferous tubules. J. Androl. 17, 615–27.CrossRefGoogle ScholarPubMed
Schlatt, S., Rosiepen, G., Weinbauer, G.F., Rolf, C., Brook, P.F. & Nieschlag, E. (1999). Germ cell transfer into rat, bovine, monkey and human testes. Human Reprod. 14, 144–50.CrossRefGoogle ScholarPubMed
Wistuba, J., Schlatt, S., Cantauw, C., von Schonfeldt, V., Nieschlag, E. & Behr, R. (2002). Transplantation of wild-type spermatogonia leads to complete spermatogenesis in testes of cyclic 3′,5′-adenosine monophosphate response element modulator-deficient mice. Biol. Reprod. 67, 1052–7.CrossRefGoogle ScholarPubMed
Yoshinaga, K., Nishikawa, S., Ogawa, M., Hayashi, S., Kunisada, T., Fujimoto, T. & Nishikawa, S. (1991). Role of c-kit in mouse spermatogenesis: identification of spermatogonia as a specific site of c-kit expression and function. Development 113, 689–99.CrossRefGoogle ScholarPubMed