载体腹腔内肝细胞移植

目的肝细胞移植可替代部分肝细胞功能及介导基因治疗。腹腔肝细胞移植操作简便，损伤小，但除脾脏、门静脉外，其他部位移植肝细胞不易存活。作者介绍一种用聚羟基乙酸－胶原载体腹腔内移植同种异体肝细胞的方法。方法将聚羟基乙酸纤维制成直径１ｃｍ的棉絮状，浸入提纯的鼠尾胶原２４小时；用胶原酶灌注收获ＳＤ大鼠单肝细胞，以８×１０／ｍｌ细胞浓度植入胶原铺底的Ｃｏｓｔａｒ　５０ｍｌ斜口瓶与聚羟基乙酸－胶原载体培养９６小时，构建聚羟基乙酸－胶原载体粘附肝细胞。移植入４０％肝切除Ｗｉｓｔａｒ大鼠的大网膜内。分别于术后２周、４周、２月处死大鼠，取移植部位病检。移植２月肝细胞用异硫氢肥分离细胞总ＲＮＡ，人工合成白蛋白，Ｐ４５０ｂ寡核苷酸探针行Ｎｏｒｔｈｅｍ杂交。结果肝细胞移植后２周至２月均在网膜移植部位发现肝细胞团，４周时已呈索状排列，肝细胞团周边可见细胞核大的新生细胞，并有分裂相。ＰＡＳ染色这些肝细胞内有糖原颗粒。这些移植入的肝细胞均有白蛋白，Ｐ４５０ｂ的ｍＲＮＡ表达。结论利用聚羟基乙酸－胶原载体粘附的肝细胞腹腔内移植易于存活并保持其特异基因表达，参与机体糖、蛋白代谢和生物转化功能。

Transplantation of hepatocytse into peritoneal cavity by network carriers made of collagen-coated polygly-colic acid fiber

Objective Intraperitoneal hepatocytes transplantation is easy of access and potentially capable of engrafting a large quantity of hepatocytes at one ted. But the efficacy of free intraperitoneal hepatocytestransplantation is controversial, for the cells are difficult to be alive and to keep their specific hepatic function.This study describes a metho attaching hepatocytes to a "polyglycolic acid-collagen carrier" in cell culture andthen implanting this polymer-cell scaffold into rat omentum. Method "Polrelycolic acid-collagen carrier" wasmade of absorbable polyglycolic acid suture and rat tail collagen. Using intra-portal vein collagenase perfusion tech technique hepatocytes from SD rats were harvested, and cocultured them with "polrelycolic acid-collagen carrier" for96 hours. Then they were implanted into omentum of Wistar rats having been partially hepatectomized. ResultSuccessful engraftment in the omentum wes observed 2 weeks, 4 weeks, 2 months after transplatation. The cellsshowed viablity and mitotic figures. PAS stain showed the implanted hepatocytes had glucogen within the cytoplasm. To assess if these implanted hepatocytes still maintianed specific hepatic gene expressions Northem blot withsynthetic oligodenoxynucleotide probes of albumin and p450b was carried out. Both albumin and P450b mRNAwere expressed in these transplanted hepatocytes. Conclusion This method of hepatocytes transplantation is al alternative to the current way and the implanted hepatocytes take part in the host's metatalic activities.