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Transplantation of isolated hepatocytes in the liver of analbuminemic rats

将分离的肝细胞移植到无蛋白血症大鼠的肝脏中

基本信息

批准号：
61570179
负责人：
OGAWA Katsuhiro
金额：
$ 1.15万
依托单位：
Asahikawa Medical College (1988)Sapporo Medical University (1986-1987)
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1986
资助国家：
日本
起止时间：
1986 至 1988
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-61570179/
关键词：
Albumin Analbuminemic Rat Isolated Hepatocytes 肝内移植

项目摘要

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.

将分离的肝细胞移植到人体部分，将适用于严重肝损伤或遗传性代谢障碍患者的治疗。尽管肝细胞已被报道可移植于脾、皮下组织、眼内等组织，但肝脏被认为是最适合移植的地方，因为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高蛋白血症大鼠的肝脏内，并通过免疫组织化学染色观察移植的肝细胞的命运。将2×106个肝细胞注入接受2/3肝部分切除的NAR门静脉。在灌流后不同时间，用抗大鼠白蛋白抗体对肝脏进行灌流固定和免疫组织化学染色。在2小时时，虽然大部分肝细胞聚集在门静脉内，但在肝窦内可见少量肝细胞。这些细胞呈近球形，与窦壁贴壁较差。移植后6h，肝细胞部分附着于宿主肝细胞，肝窦内皮细胞的胞浆突起部分缺失。12h时，移植细胞已完全整合到宿主肝索内并在宿主肝组织内组织起来。结果表明，门静脉注入的肝细胞很容易在宿主肝脏内结合和组织起来。