Obara T, Conti C J, Baba M, Resau J H, Trifillis A L, Trump B F, Klein-Szanto A J
Am J Pathol. 1986 Mar;122(3):386-91.
A rapid and sensitive method for the identification of human tissues xenotransplanted in nude mice was developed. An in situ hybridization technique made it possible to distinguish between cells of human origin and cells of murine origin in formalin-fixed paraffin sections. High-molecular-weight DNAs extracted from human or mouse tissues were sonicated, nick-translated with 32P-dCTP, and used as hybridization probes. Dot blot hybridization of 32P-labeled probes revealed clear species-specific signals. Formalin-fixed paraffin-embedded tissue samples from repopulated tracheal transplants, containing either human tracheal epithelial cells or human renal tubular cells, were used. Cells of human and murine origin were distinguishable by in situ hybridization with sonicated DNA probes. This method has several advantages; simple preparation of probes, high sensitivity, and applicability to formalin-fixed paraffin-embedded tissue sections. In situ hybridization with sonicated DNA probes should provide a powerful tool for verifying the human origin of xenotransplanted tissues in nude mice.
开发了一种快速、灵敏的方法用于鉴定裸鼠体内移植的人体组织。原位杂交技术能够在福尔马林固定的石蜡切片中区分人源细胞和鼠源细胞。从人或小鼠组织中提取的高分子量DNA经超声处理后，用32P-dCTP进行缺口平移，并用作杂交探针。32P标记探针的斑点印迹杂交显示出清晰的物种特异性信号。使用了来自重新植入的气管移植组织的福尔马林固定石蜡包埋组织样本，这些样本包含人气管上皮细胞或人肾小管细胞。通过用超声处理的DNA探针进行原位杂交，可以区分人源和鼠源细胞。该方法具有几个优点：探针制备简单、灵敏度高以及适用于福尔马林固定的石蜡包埋组织切片。用超声处理的DNA探针进行原位杂交应为验证裸鼠体内异种移植组织的人源性质提供一个有力工具。
