Transplantation of isolated hepatocytes in the liver of analbuminemic rats

局部分离肝细胞移植将适用于严重肝损伤或遗传性代谢紊乱患者的治疗。虽然有报道称肝细胞可移植于脾脏、皮下组织、眼内间隙等，但肝脏被认为是最适合移植的地方，因为肝门静脉血流中存在的肝营养因子将为肝细胞提供合适的环境。在本项目中，我们将正常肝细胞移植到无抗体血症大鼠的肝脏内，并通过大鼠白蛋白免疫组化染色研究移植肝细胞的命运。采用胶原酶灌注法从SB大鼠肝脏中分离肝细胞。然后将2 × 10^6个肝细胞输注于2/3肝部分切除的NAR门静脉。灌注后不同时期，肝脏灌注固定，用抗大鼠白蛋白抗体免疫染色。在2h时，虽然大部分注入的肝细胞聚集在门静脉内，但在窦内观察到少量肝细胞。这些细胞几乎呈球形，与窦壁的附着性很差。6h时，移植肝细胞部分附着在宿主肝细胞上，其中窦状内皮细胞的细胞质突起部分缺失。12h时，移植细胞完全融入宿主肝索内，并在宿主肝组织内组织。结果表明，经门静脉输注的肝细胞在宿主肝脏内易于整合和组织。

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.