정원줄기세포의 이종간 이식을 위한 효율적인 수여(Recipient) 생쥐 생산 기법 개발

초록

Spermatogonial xenotransplantation assay provides access to the several species germline and has been used in experimental animal models to study stem cell biology and germline development. To make the xenotransplantation technology more extensively accessible, it is require development of recipient preparation protocols. Therefore, we propose an optimal method for preparation of athymic nude recipient with busulfan treatment. To investigate the optimal concentration of busulfan treatment, athymic nude mice (6 similar to 8 weeks old) were intraperitoneally injected with different concentrations of busulfan (0, 15, 25, 35 and 45 mg/kg). 6 similar to 8 weeks after busulfan treatment, testicular weight was significantly lower in 45 mg/kg treated group than the other groups. And the percentage of spermatogenic tubule was significantly reduced in 45 mg/kg of busulfan treated recipient testis. Also we compared the gene expression level of sertoli-cell-derived growth factor, glial-cell-line-derived neurotrophic factor (GDNF) as a potential measure of niche function. Relative to the other experimental groups, 45 mg/kg busulfan treated recipient testis showed higher GDNF expression. Next, to find out the appropriate time to transplantation after treated with busulfan (45 mg/kg), the rat testicular donor cells were transplanted into recipient testis at 2, 4, 6 and 8 weeks after treated with busulfan. Analysis of recipient mice revealed that transplantation of donor cells on 6 weeks after busulfan treatment was the most effective time to support high level of donor stem cell engraftment. In this study, we provide an optimum experimental condition for preparation of recipient mouse model system for xenotransplantation. Also, our data could contribute to recipient preparation in other species.

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