Ye J, Wang H M, Ogden T E, Ryan S J
Department of Ophthalmology, University of Southern California School of Medicine, Los Angeles.
Curr Eye Res. 1993 Jul;12(7):629-39. doi: 10.3109/02713689309001842.
To facilitate identification of transplanted retinal pigment epithelial (RPE) cells, we sought to double-label the cells with 5-bromodeoxyuridine (BrdU) and with natural pigment. The BrdU is not lost during cell division but does require immunohistochemical methods for visualization; the pigment, on the other hand, allows immediate, obvious identification, but is gradually lost with cell division. Together they provide a convenient, long-term double label. Herein we report the successful allotransplantation of double-labelled RPE cells onto Bruch's membrane of albino rabbits. The labelled cells were localized by anti-BrdU antibody and the avidin biotin-alkaline phosphatase complex (ABC-AP) method, and by visible inclusions of pigment. Using this double-label method, allotransplanted RPE cells could be readily and reliably identified on the recipient Bruch's membrane eight months after transplantation. The cells had distinct basal and apical morphology, and were in close contact with the photoreceptor outer segments of the host. This successful allotransplantation raises the possibility that the subretinal space of the rabbit might enjoy some degree of immunologic privilege.
为便于识别移植的视网膜色素上皮（RPE）细胞，我们尝试用5-溴脱氧尿苷（BrdU）和天然色素对细胞进行双重标记。BrdU在细胞分裂过程中不会丢失，但需要通过免疫组织化学方法才能观察到；另一方面，色素能立即实现明显的识别，但会随着细胞分裂逐渐丢失。二者共同提供了一种便捷的长期双重标记方法。在此，我们报告了将双重标记的RPE细胞成功同种异体移植到白化兔布鲁赫膜上的情况。通过抗BrdU抗体和抗生物素蛋白-生物素-碱性磷酸酶复合物（ABC-AP）方法，以及色素的可见内含物来定位标记细胞。使用这种双重标记方法，在移植八个月后，可在受体的布鲁赫膜上轻松且可靠地识别出同种异体移植的RPE细胞。这些细胞具有明显的基底和顶端形态，并与宿主的光感受器外段紧密接触。这种成功的同种异体移植增加了兔子视网膜下间隙可能享有一定程度免疫赦免的可能性。
