Transplantation of isolated hepatocytes in the liver of analbuminemic rats

离体肝细胞移植将适用于严重肝损伤或遗传代谢紊乱患者的治疗。虽然肝细胞已被报道可移植到脾、皮下组织、眼内空间等，肝脏被认为是最适合移植的地方，因为存在于门静脉流中的肝营养因子将为肝细胞提供合适的环境。本研究将正常肝细胞移植到无白蛋白血症大鼠的肝内，通过免疫组化方法观察移植肝细胞的转归，并采用胶原酶灌流法分离SB大鼠的肝细胞。然后将2 x 10^6个肝细胞输注到已接受2/3部分肝切除的NAR的门静脉中。在输注后的不同时期，肝脏灌注固定和免疫染色使用抗大鼠白蛋白抗体。在2 h，虽然大多数输注的肝细胞聚集在门静脉内，少量肝细胞内观察到的窦。这些细胞的形状接近球形，与窦壁的附着较差。6 h时，移植肝细胞部分贴附于宿主肝细胞，肝窦内皮细胞胞质突起部分缺失。12 h时，移植肝细胞完全整合于宿主肝索内，并在宿主肝组织内组织化，结果表明门静脉输注的肝细胞易于整合于宿主肝组织内。

Transplantation of isolated hepatocytes in the part of the body will be applicable to the therapy of patients with severe liver damage or genetic metabolic disorders. Although the hepatocytes have been reported to be transplantable in the spleen, subcutaneous tissue, intraocular space, etc., the liver is considered to be the most suitable place for transplantation because hepatotrophic factors present in the portal flow will provide appropriate environment for the hepatocytes. In this project, we transplanted normal hepatocytes within the liver of analbuminemic rats and investigated the fate of transplanted hepatocytes by immunohistochemical staining of rat albumin.Hepatocytes were isolated from the liver of SB rats by collagenase perfusion method. Then 2 x 10^6 hepatocytes were infused into the portal vein of NAR which had received 2/3 partial hepatectomy. Various periods after infusion, the liver was perfusion-fixed and immunostained using an anti rat-albumin antibody.At 2h, although the most infused hepatocytes were aggregated within the portal vein, a small number of hepatocytes were observed within the sinusoids. These cells were nearly spherical in shape and poorly attached to the sinusoidal wall. At 6h, the transplanted hepatocytes partly attaached to the host hepatocytes where the cytoplasmic processes of the sinusoidal endothelial cells were partly deleted. At 12h, the transplanted cells were completely incorporated within the host hepatic cords and organized within the host hepatic tissue.The results showed that the hepatocytes infused in the portal vein are readily incorporated and organized within the host liver.