Successful transplantation of bovine testicular cells to heterologous recipients
Successful transplantation of bovine testicular cells to heterologous recipients
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DOI：
10.1530/rep.1.01125
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发表时间：
2006-10-01
期刊：
影响因子：
3.8
通讯作者：
Hill, Jonathan R.
中科院分区：
文献类型：
--
作者：
Herrid, Muren;Vignarajan, Soma;Hill, Jonathan R.
虽然异种生殖细胞移植在猪和山羊身上取得了成功，但据报道，仅自体移植就能在牛身上导致供者来源的精子发生。本研究的目的是探讨异种生殖细胞移植是否会导致受体睾丸在不同品种的牛身上定植。从8头金牛供体公犊中分离出睾丸细胞，然后将其移植到15头牛指数杂交公犊中。所有动物均为青春期前，供体年龄为5~7个月，受体年龄为5~11个月，阴囊周长15~22 cm。将酶消化法制备的供体睾丸细胞单细胞悬液，用荧光染料PKH26或CFDA-SE标记后，在超声引导下移植到受体睾丸网状结构。为了评估供体细胞的定植寿命，受者在细胞移植后2-30周被去势。在接受PKH26标记细胞的睾丸中，有15/25(60%)观察到供体细胞，而在任何受体中都没有发现CFDA-SE阳性细胞。捐赠者或接受者的成熟度(以阴囊周长衡量)不影响殖民潜力。在新分离的小管中，观察到PKH26阳性细胞团，这表明细胞分裂或广泛的局部定植于特定区域的小管。冰冻切片中，在曲细精管基底膜上可见PKH26阳性细胞，提示这些细胞已成功地从曲细管腔内迁移出来，可能是精原细胞。我们认为，PKH26更适合于标记供体睾丸细胞，移植后6个月即可鉴定出供体细胞。这些结果表明，同种异体睾丸细胞可以在金牛和指数牛之间进行移植。进一步的研究将调查移植的睾丸细胞的功能。
While heterologous germ cell transplantation was successful in pigs and goats, autologous transplantation alone has been reported to result in donor-derived spermatogenesis in cattle. The objective of this study was to investigate whether the transplantation of heterologous germ cells could result in colonization of recipient testes in cattle of different breeds. Testicular cells were isolated from 8 Bos taurus donor bull calves and then transferred into 15 Bos indices-cross bull calves. All animals were pre-pubertal, donors were aged 5-7 months and recipients 5-11 months, and scrotal circumferences ranged from 15 to 22 cm. Single cell suspensions of donor testicular cells, prepared by enzymatic digestion, were labelled with fluorescent dyes PKH26 or CFDA-SE, before transfer into the rete testis of recipients under ultrasonographic guidance. To assess the longevity of colonization by donor cells, recipients were castrated 2-30 weeks after cell transfer. Donor cells were observed in 15/25 (60%) of the testes that received PKH26-labelled cells, whereas no CFDA-SE-positive cell was identified in any recipients. The maturity of the donors or recipients (measured by scrotal circumference) did not affect colonization potential. In freshly isolated tubules, clumps of PKH26-positive cells were observed, which indicated either cell division or extensive local colonization of specific areas of the tubules. In frozen sections, PKH26-positive cells were identified on the seminiferous tubule basement membrane, which indicated that these cells had successfully migrated from the tubule lumen and were likely to be spermatogonia. We conclude that PKH26 was more suitable for labelling donor testis cells and donor cells can be identified up to 6 months following transfer. These results indicate that allogeneic transplantation of testicular cells can occur between Bos taurus and Bos indices cattle. Further studies will investigate functionality of transferred testicular cells.