骨髓源性肝干细胞定向分化及脾内移植研究

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1、维普资讯http://www.cqvip.com中华实验外科杂志2004年5月第21卷第5期ChinJExpSurg，May2004，Vol21，No．5·551··实验研究·骨髓源性肝干细胞定向分化及脾内移植研究蔡云峰陈积圣闭军魏菁何劲松莫隽全【摘要】目的寻找骨髓源性肝干细胞的表面标记，进行定向分化及脾内移植研究。方法流式细胞仪检测淤胆大鼠骨髓中干细胞群体的数量变化，寻找肝干细胞标记，免疫磁珠分离，进行体外诱导分化和肝再生模型的脾内移植，观察细胞形态的变化，免疫组织化学和免疫荧光技术检测白蛋白、AFP、CK8／18等肝细胞标记的表达

2、。结果淤胆鼠B2微球蛋白阴性(m一)细胞数量较对照组数量明显增高，分别为(6．17±2．70)％和(O．79±0．61)％(P<0．01)，m一细胞体外培养及脾内移植均可出现肝细胞样细胞，白蛋白、AFP、CK8／18表达阳性。结论D2m一细胞在体内外具有向肝细胞分化的能力，脾内移植是肝干细胞移植可供选择的部位之一。【关键词】骨髓干细胞；肝细胞；分化；移植Differentiationinductionandintrasplenicaltransplantationofbonemarrow·derivedliverstemcellsCA

3、IYun—feng，C艘NJi—sheng，MTN，eta1．DepartmentofHepaticandBiliarySurgery，PSec—ondA所liatedHospitalofSunYat—senUniversity，Guangzhou510120，China【Abstract】0bjectiveToidentifythesurfacemarkerofbonemarrow—derivedliverstemcells，iso—latethestemcells，andinvestigatethedifferentiationa

4、ndintrasplenicaltransplantationofthestemcells．MethodsThevariationsofseveralstemcellsurfacemarkers，including82一microglobulinnegative(Ibm一)，Thy一1，CD34，F1t一3，IL一3R，andc—kitmarkers，weredetectedinthebonemarowofcholestaticratsbyusingflowcytometry．Thesignificantcellpopulations

5、(m—cells)werethenisolatedusingamagneticbeadcell—sortingprocedure．Theisolatedcellswereculturedinasystemcontainingcholestaticserumandhepatocytegrowthfactor(HGF)．Theywerealsotransplantedintothespleenofaratmode1．whichhadbeentreatedwithretrorsineand70％partialhepatectomy．Them

6、orphologyofthetellswasobserved，andtheexpressionsofalbumin，AFP，andCK8／18wasdetectedbyusingimmunohis—tochemistryandimmunofluorescense．Resultsm—cellswere(0．79±0．61)％inthebonemarrowofnormalrats，significantlylessthaninthecholestaticrats[(6．17±2．70)％，P<0．01]．Afterbeingco-cult

7、uredwithcholestaticserumandHGF，thecellsshowedmultilateraltransformationandresembledhep—atcytesmorphologically．Inthespleenofthemodelrats，hepatocyte—likeceilsappeared．Boththeculturedcellsandthetransplantedcellsexpressedalbumin，AFP，andCK8／18，allknowntobethecharacteristicma

8、rkersofhepatocyte．Conclusionm—cellshavetheabilitytotrans—differentiatedintohepatocytebothinvivoandinvitro．Sple