Theise N D, Nimmakayalu M, Gardner R, Illei P B, Morgan G, Teperman L, Henegariu O, Krause D S
Department of Pathology, New York University, School of Medicine, New York, NY 10016, USA.
Hepatology. 2000 Jul;32(1):11-6. doi: 10.1053/jhep.2000.9124.
It has been shown in animal models that hepatocytes and cholangiocytes can derive from bone marrow cells. We have investigated whether such a process occurs in humans. Archival autopsy and biopsy liver specimens were obtained from 2 female recipients of therapeutic bone marrow transplantations with male donors and from 4 male recipients of orthotopic liver transplantations from female donors. Immunohistochemical staining with monoclonal antibody CAM5.2, specific for cytokeratins 8, 18, and 19, gave typical strong staining of hepatocytes, cholangiocytes, and ductular reactions in all tissues, to the exclusion of all nonepithelial cells. Slides were systematically photographed and then restained by fluorescence in situ hybridization (FISH) for X and Y chromosomes. Using morphologic criteria, field-by-field comparison of the fluorescent images with the prior photomicrographs, and persistence of the diaminiobenzidene (DAB) stain through the FISH protease digestion, Y-positive hepatocytes and cholangiocytes could be identified in male control liver tissue and in all study specimens. Cell counts were adjusted based on the number of Y-positive cells in the male control liver to correct for partial sampling of nuclei in the 3-micron thin tissue sections. Adjusted Y-positive hepatocyte and cholangiocyte engraftment ranged from 4% to 43% and from 4% to 38%, respectively, in study specimens, with the peak values being found in a case of fibrosing cholestatic recurrent hepatitis C in one of the liver transplant recipients. We therefore show that in humans, hepatocytes and cholangiocytes can be derived from extrahepatic circulating stem cells, probably of bone marrow origin, and such "transdifferentiation can replenish large numbers of hepatic parenchymal cells.
动物模型研究表明，肝细胞和胆管细胞可源自骨髓细胞。我们研究了这一过程是否发生在人类身上。从2名接受男性供体治疗性骨髓移植的女性受者以及4名接受女性供体原位肝移植的男性受者身上获取了存档的尸检和活检肝脏标本。使用对细胞角蛋白8、18和19具有特异性的单克隆抗体CAM5.2进行免疫组织化学染色，所有组织中的肝细胞、胆管细胞和小胆管反应均呈现典型的强染色，排除了所有非上皮细胞。对玻片进行系统拍照，然后通过荧光原位杂交（FISH）对X和Y染色体进行重新染色。利用形态学标准，逐视野将荧光图像与先前的显微照片进行比较，并通过FISH蛋白酶消化使二氨基联苯胺（DAB）染色持续存在，从而在男性对照肝组织和所有研究标本中鉴定出Y阳性的肝细胞和胆管细胞。根据男性对照肝中Y阳性细胞的数量调整细胞计数，以校正3微米薄组织切片中细胞核的部分取样。在研究标本中，调整后的Y阳性肝细胞和胆管细胞植入率分别为4%至43%和4%至38%，其中一名肝移植受者的纤维化胆汁淤积性复发性丙型肝炎病例中出现了峰值。因此，我们证明在人类中，肝细胞和胆管细胞可源自肝外循环干细胞，可能起源于骨髓，并且这种“转分化”可以补充大量肝实质细胞。
