CAJ | 학술논문

探讨 CM-DiI 标记大鼠原代肝细胞的最佳浓度、细胞的移植量和移植途径．采用原位胶原酶灌注和密度梯度离心法分离、纯化大鼠肝细胞，用不同浓度 CM-DiI 进行标记，筛选出 CM-DiI 的最佳标记浓度，分别从门静脉、尾静脉移植到2/3肝切除0 h 大鼠，对细胞移植36 h 后的肝组织石蜡切片和冰冻切片分别进行免疫组织化学分析和荧光观察．结果显示，4μmol?mL -1浓度的 CM-DiI 和肝细胞孵育5 min ，细胞标记率高达95％．每克大鼠残肝移植0.8 mL的细胞悬液（107个?mL -1）和0.2，0.4，0.6 mL 移植后检测到的标记细胞数目相比有显著性差异（P＜0.01），经肝门静脉移植后可以检测到的标记细胞数比尾静脉多（P＜0.01）．CM-DiI 标记大鼠原代肝细胞的最佳浓度为4μmol?mL -1，标记细胞的最佳移植量为每克大鼠残肝移植0.8 mL 的细胞悬液（107个?mL -1）、最佳移植途径为经肝门静脉移植．
탐토 CM-DiI 표기대서원대간세포적최가농도、세포적이식량화이식도경．채용원위효원매관주화밀도제도리심법분리、순화대서간세포，용불동농도 CM-DiI 진행표기，사선출 CM-DiI 적최가표기농도，분별종문정맥、미정맥이식도2/3간절제0 h 대서，대세포이식36 h 후적간조직석사절편화빙동절편분별진행면역조직화학분석화형광관찰．결과현시，4μmol?mL -1농도적 CM-DiI 화간세포부육5 min ，세포표기솔고체95％．매극대서잔간이식0.8 mL적세포현액（107개?mL -1）화0.2，0.4，0.6 mL 이식후검측도적표기세포수목상비유현저성차이（P＜0.01），경간문정맥이식후가이검측도적표기세포수비미정맥다（P＜0.01）．CM-DiI 표기대서원대간세포적최가농도위4μmol?mL -1，표기세포적최가이식량위매극대서잔간이식0.8 mL 적세포현액（107개?mL -1）、최가이식도경위경간문정맥이식．
The optimum concentration of CM-DiI incubated with primary hepatocyte of rat , optimum transplant volume and route of labeled primary hepatocyte are studied . The rat hepatocytes are isolated and purified by collagenase and density gradient centrifugation , and labeled with different concentration of CM-DiI , and then transplanted on rat which are previously suffered 2/3 partial hepatectomy 0 h by liver portal vein and tail vein , respectively . The livers are removed after transplanting hepatocyte 36 h , frozen section and paraffin section are prepared and analyzed by immunohistochemistry and fluorescence observation , respectively . The results show that hepatocyte can be labeled to 95% after incubating with CM-DiI 4 μmol?mL - 1 for 5 min , labeled hepatocytes which are observed on frozen section transplanted by 0.8 mL per keilogram remnant liver weight are more than that of 0.2 ,0.4 and 0.6 mL (P < 0.01) , and labeled hepatocytes transplanted by liver portal vein are more than that of tail vein ( P < 0.01 ) . Hepatocytes incubated with CM-DiI 4 μmol?mL - 1 for 5 min can be labeled to 95% , and 0.8 mL labeled hepatocytes per keilogram remnant liver weight transplanted by liver portal vein can get preferable label and transplantation efficiency .