Two allogeneic descendents derived from the high-dose busulfan-treated infertile mouse model after freeze-thawed spermatogonial stem cell transplantation.
OBJECTIVE
To evaluate restoration of spermatogenesis and fertility of the recipients who underwent high-dose ablative treatment before freeze-thawed spermatogonial stem cell (SSC) allotransplantation.


METHODS
Prospective experimental study.


METHODS
University-based teaching hospital.


METHODS
Adult busulfan-treated BALB/c nude mice as recipients of C(57)BL/6 pup SSCs, oocytes of 4-to 6-week-old F(1) hybrid mice for IVF, adult closed group ICR mouse as pseudopregnant female.


METHODS
Isolation, purification, and fresh and freeze-thawed transplantation, IVF, embryo transplantation.


METHODS
Cell viability, Western blot, real-time fluorescence quantitative-polymerase chain reaction (PCR), scanning electron microscope (SEM), outcome of IVF, and embryo transplantation.


RESULTS
Western blot, real-time fluorescence quantitative-PCR, and SEM revealed different spermatogenesis where the fresh groups were uniquely higher than the freeze-thawed groups. A total of six developed two-cell embryos obtained by IVF were transplanted into the oviduct of a pseudopregnant ICR mouse, which resulted in a birth of two paternally different pups. Two female offspring grew into healthy adults with no apparent abnormalities.


CONCLUSIONS
The results provide the first solid evidence that spermatozoa generated from the busulfan-treated recipient and donor cryopreserved SSCs were both fertile. Ultimate evaluation of the SSC cryopreservation and transplantation technology rests on their functional capacity to generate desirable donor spermatogenesis and fertility in the recipient.
Wang
Xiang
X
Department of Urology, Huashan Hospital, Fudan University, Shanghai, P. R. China. seanw@medmail.com.cn
Ding
Qiang
Q
Zhang
Yuanfang
Y
Wang
Huilin
H
Ma
Lianghong
L
Xie
Xiayang
X
D000818:Animals; D002066:Busulfan / Q000009:adverse effects*; D002454:Cell Differentiation; D049109:Cell Proliferation; D002470:Cell Survival; D015925:Cryopreservation; D005260:Female; D005307:Fertilization in Vitro; D006801:Humans; D007248:Infertility, Male / Q000503:physiopathology / Q000628:therapy*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008813:Mice, Inbred ICR; D008819:Mice, Nude; D013091:Spermatogenesis; D013094:Spermatozoa / Q000502:physiology / Q000637:transplantation*; D033581:Stem Cell Transplantation; D014183:Transplantation, Heterologous
D002066:Busulfan