大连医科大学学报
大連醫科大學學報
대련의과대학학보
JOURNAL OF DALIAN MEDICAL UNIVERSITY
2013年
6期
538-542
,共5页
张春兴%王芳%廖彩仙%吴亚琼%蒋建武
張春興%王芳%廖綵仙%吳亞瓊%蔣建武
장춘흥%왕방%료채선%오아경%장건무
骨髓衍生肝干细胞%免疫磁珠分选%体外扩增培养%干细胞移植%c-Kit+Lin-细胞
骨髓衍生肝榦細胞%免疫磁珠分選%體外擴增培養%榦細胞移植%c-Kit+Lin-細胞
골수연생간간세포%면역자주분선%체외확증배양%간세포이식%c-Kit+Lin-세포
bone marrow derived liver stem cells%MACS%expand and culture in vitro%stem cell transplantation%c-Kit+Lin-cells
目的：研究移植刚分选及扩增后雄性小鼠的骨髓c-Kit＋Lin-细胞到酒精性肝纤维化雌性模型小鼠体内后，模型小鼠的肝脏形态改变、肝功能变化和移植细胞的分布、分化及归巢情况。方法采用酒精灌胃法建立酒精性肝纤维化雌性小鼠模型，造模成功后随机选取8只模型小鼠（模型组）与正常小鼠8只（对照组）比较两组肝功能变化。剩余模型小鼠再随机选取32只，分为4组处理，并再随机选取8只正常小鼠（第1组）与其对照。模型小鼠的4组分别为：移植前取血处死组（第2组），移植新鲜分选的c-Kit＋Lin-细胞组（第3组），移植扩增后的c-Kit＋Lin-细胞组（第4组），仅予以注射Buffer组（第5组）。采用免疫磁珠法分选雄性小鼠骨髓c-Kit＋Lin-细胞；利用SCF＋HGF＋FL＋LIF ＋TPO＋IL-3因子组合对其进行体外扩增；将刚分选及扩增后c-Kit＋Lin-细胞移植入酒精性肝纤维化雌性模型小鼠体内，检测两组肝功能改变、肝脏纤维化改善、含雄性小鼠Y染色体性别决定基因cDNA（Sry）的移植细胞在受体肝内定居分化情况。结果（1）两移植组在移植30天后，较移植前肝脏纤维化改善明显；（2）两移植组在移植30天后AST、ALT、ALB与移植前比较差异有显著性意义（P＜0．05）；（3）两移植组均可见含雄性小鼠Y染色体性别决定基因cDNA（ Sry）的移植细胞，并同时表达ALB mRNA，计数细胞比例，比较差异无显著性意义（P＞0．05）。结论移植c-Kit＋Lin-细胞后能在受体肝内定居并转化成有功能新生肝细胞；移植c-Kit＋Lin-细胞后能明显改善肝损伤小鼠的肝功能及肝脏纤维化；扩增对c-Kit＋Lin-细胞的归巢、在体内分化没有明显影响。
目的：研究移植剛分選及擴增後雄性小鼠的骨髓c-Kit＋Lin-細胞到酒精性肝纖維化雌性模型小鼠體內後，模型小鼠的肝髒形態改變、肝功能變化和移植細胞的分佈、分化及歸巢情況。方法採用酒精灌胃法建立酒精性肝纖維化雌性小鼠模型，造模成功後隨機選取8隻模型小鼠（模型組）與正常小鼠8隻（對照組）比較兩組肝功能變化。剩餘模型小鼠再隨機選取32隻，分為4組處理，併再隨機選取8隻正常小鼠（第1組）與其對照。模型小鼠的4組分彆為：移植前取血處死組（第2組），移植新鮮分選的c-Kit＋Lin-細胞組（第3組），移植擴增後的c-Kit＋Lin-細胞組（第4組），僅予以註射Buffer組（第5組）。採用免疫磁珠法分選雄性小鼠骨髓c-Kit＋Lin-細胞；利用SCF＋HGF＋FL＋LIF ＋TPO＋IL-3因子組閤對其進行體外擴增；將剛分選及擴增後c-Kit＋Lin-細胞移植入酒精性肝纖維化雌性模型小鼠體內，檢測兩組肝功能改變、肝髒纖維化改善、含雄性小鼠Y染色體性彆決定基因cDNA（Sry）的移植細胞在受體肝內定居分化情況。結果（1）兩移植組在移植30天後，較移植前肝髒纖維化改善明顯；（2）兩移植組在移植30天後AST、ALT、ALB與移植前比較差異有顯著性意義（P＜0．05）；（3）兩移植組均可見含雄性小鼠Y染色體性彆決定基因cDNA（ Sry）的移植細胞，併同時錶達ALB mRNA，計數細胞比例，比較差異無顯著性意義（P＞0．05）。結論移植c-Kit＋Lin-細胞後能在受體肝內定居併轉化成有功能新生肝細胞；移植c-Kit＋Lin-細胞後能明顯改善肝損傷小鼠的肝功能及肝髒纖維化；擴增對c-Kit＋Lin-細胞的歸巢、在體內分化沒有明顯影響。
목적：연구이식강분선급확증후웅성소서적골수c-Kit＋Lin-세포도주정성간섬유화자성모형소서체내후，모형소서적간장형태개변、간공능변화화이식세포적분포、분화급귀소정황。방법채용주정관위법건립주정성간섬유화자성소서모형，조모성공후수궤선취8지모형소서（모형조）여정상소서8지（대조조）비교량조간공능변화。잉여모형소서재수궤선취32지，분위4조처리，병재수궤선취8지정상소서（제1조）여기대조。모형소서적4조분별위：이식전취혈처사조（제2조），이식신선분선적c-Kit＋Lin-세포조（제3조），이식확증후적c-Kit＋Lin-세포조（제4조），부여이주사Buffer조（제5조）。채용면역자주법분선웅성소서골수c-Kit＋Lin-세포；이용SCF＋HGF＋FL＋LIF ＋TPO＋IL-3인자조합대기진행체외확증；장강분선급확증후c-Kit＋Lin-세포이식입주정성간섬유화자성모형소서체내，검측량조간공능개변、간장섬유화개선、함웅성소서Y염색체성별결정기인cDNA（Sry）적이식세포재수체간내정거분화정황。결과（1）량이식조재이식30천후，교이식전간장섬유화개선명현；（2）량이식조재이식30천후AST、ALT、ALB여이식전비교차이유현저성의의（P＜0．05）；（3）량이식조균가견함웅성소서Y염색체성별결정기인cDNA（ Sry）적이식세포，병동시표체ALB mRNA，계수세포비례，비교차이무현저성의의（P＞0．05）。결론이식c-Kit＋Lin-세포후능재수체간내정거병전화성유공능신생간세포；이식c-Kit＋Lin-세포후능명현개선간손상소서적간공능급간장섬유화；확증대c-Kit＋Lin-세포적귀소、재체내분화몰유명현영향。
Objective To elucidate the liver function recovery and the translated stem cells ’ location and differentiation of the hepatic fibrosis female rats induced by alcohol in the group of translated expanded and fresh isolated c -Kit+Lin-cells.Methods We used alcohol to establish the female rats hepatic fibrosis model .After the modeling randomly selected eight mice (model group) compared with normal mice (control group) liver function.The remaining mice were randomly selected 32 divided into 4 groups.Randomly selected eight normal mice (group 1)as a control group.C-kit+Lin-cells were isolated from mouse bone marrow by using a high -gradient magnetic cell sorting system ( MACS) ,and expanded with combinations of SCF +HGF+FL+LIF +TPO+IL-3 for 7days in a liquid culture system .We translated the expanding c-kit+Lin-cells into the normal female rats and hepatic fibrosis female rats induced by alcohol .Observe the translated stem cells'location and differentiation in the rats body and count the cells which expressed both gene of Sry , and compare the two group.Results (1) After 30 days,HE and VG histological examination showed that the translated groups were less fibrosis and the control group obviously .(2) After 30 days,AST,ALT,ALB after transplantation compared with pretrans-plant significant difference (P<0.05).(3) Results of hybridization of Y -chromosome determined cDNA and albumin mRNA indicated that in the translated group there existed hepatic cells which derived from stem cells in the liver .The difference was not statistically significant (P>0.05).Conclusion In this experiment condition,expanding c -Kit+Lin-cell did not effect the stem cells location and differentiation .The liver function recovery in the group translated and fresh i-solated c-Kit+Lin-cells ,reverse the liver fibrosis and there existed hepatic cells which derived from stem cells in the liv-er.
