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Germ Cell Transplantation in an Azoospermic Klinefelter Bull¹

Institute of Animal Sciences,³ Swiss Federal Institute of Technology Zurich, CH-8092 Zurich, Switzerland Department of Farm Animals⁴ Institute of Zoology,⁵ University of Zurich, CH-8057 Zurich, Switzerland Institute of Reproductive Medicine,⁶ University of Munster, D-48149 Munster, Germany Department of Animal Husbandry,⁷ Chulalongkorn University, Bangkok 10330, Thailand

Germ cell transplantation is a technique that transfers donor testicular cells into recipient testes. A population of germ cells can colonize the recipient testis, initiate spermatogenesis, and produce sperm capable of fertilization. In the present study, a nonmosaic Klinefelter bull was used as a germ cell recipient. The donor cell suspension was introduced into the rete testis using ultrasound-guided puncture. A pulsatile administration of GnRH was performed to stimulate spermatogenesis. The molecular approach to detect donor cells was done by a quantitative polymerase chain reaction with allele discrimination based on a genetic mutation between donor and recipient. Therefore, a known genetic mutation, associated with coat-color phenotype, was used to calculate the ratio of donor to recipient cells in the biopsy specimens and ejaculates for 10 mo. After slaughtering, meiotic preparations were performed. The injected germ cells did not undergo spermatogenesis. Six months after germ cell transplantation, the donor cells were rejected, which indicates that the donor cells could not incorporate in the testis. The hormone stimulation showed that the testosterone-producing Leydig cells were functionally intact. Despite subfertility therapy, neither the recipient nor the donor cells underwent spermatogenesis. Therefore, nonmosaic Klinefelter bulls are not suitable as germ cell recipients. Future germ cell recipients in cattle could be mosaic Klinefelters, interspecies hybrids, bulls with Sertoli cell-only syndrome, or bulls with disrupted germ cell migration caused by RNA interference.

¹ Supported by grants from the Swiss Federal Institute of Technology Zurich (ETH Project 0-20944-01, ETH-Project 0-20994-02).

² Correspondence: Hannes Joerg, Institute of Animal Sciences, ETH Zurich, Tannenstr. 1, CH-8092 Zurich, Switzerland. FAX: 41 1 632 11 67; hannes.joerg{at}inw.agrl.ethz.ch

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