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1、大鼠骨髓间充质干细胞体外诱导向神经细胞样细胞分化[摘要]目的研究大鼠骨髓间充质干细胞(BMSCs)离体分离和培养方法,探讨碱性成纤维细胞生长因子(bFGF)、表皮生长因子(EGF)、全反式维甲酸(RA)、骨形态发生蛋白-7(BMP-7)在体外诱导BMSCs向神经干细胞(NSCs)分化的作用。方法采用出生4周SD大鼠的全骨髓细胞进行培养,传至第3代时,分为3组:A组,bFGF+EGF+RA进行诱导分化;B组,BMP-7进行诱导分化;C组,不加因子继续培养。在倒置显微镜下每日观察、记录BMSCs的诱导分化情况,并应用免疫组化技术对细胞进行兔抗神经元特异性烯醇化酶(N
2、SE)单抗鉴定。结果A组诱导7d后有部分细胞具备神经元样细胞形态,胞体呈锥形或圆形,有较长单极或多极的突起,有NSE阳性细胞表达。而B组、C组细胞仍为典型的成纤维细胞形态,无NSE阳性细胞。结论bFGF、EGF和RA可以联合诱导BMSCs向NSCs分化,并可调控神经元分化,而BMP-7在此种诱导条件下未见明显的诱导BMSCs向NSCs分化的作用。�[关键词]骨髓间充质干细胞;神经干细胞;成纤维细胞生长因子;表皮生长因子;全反式维甲酸;骨形态发生蛋白-7�[中图分类号]R329.21;R329.24[文献标识码]A12[文章编号]1671-7562(2008)02
3、-0102-04��Studyonthecultureanddifferentiationofbone�marrowstromalcellsfromadultrats��YUHai-liu1,JIANGZan-li�2,MAOZu-bin�2(1.SchoolofClinicalMedicalScience,SoutheastUniversity,Nanjing210009,China;2.Department�ofOrthopaedics,ZhongdaHospital,SoutheastUniversity,Nanjing210009,China)��Abs
4、tract:ObjectiveToinvestigatetheisolatingandculturingmethodsofratbonemarrowstromalcells(BMSCs)invitroandtostudywhetheradultBMSCscouldbeinducedtodifferentiateintoneuralstemcells(NSCs)andneuronsbybasicfibroblastgrowthfactor(bFGF),epidermalgrowthfactor(EGF),all-transretinoicacid(RA)andbo
5、nemorphogeneticprotein-7(BMP-7).MethodsAdultmarrowstromalcellswereculturedfrombonemarrowoffourweek-oldSDrats.bFGF+EGF+RA(groupA)andBMP-7(groupB)wereusedtoinducethethird-generationBMSCs,andcontrastedwiththecellsthathadnotbeeninduced.Phasecontrast�microscopewasusedtoobservethe12differe
6、ntiationofBMSCsdaily.ImmunohistochemistrytechniquewasusedtoidentifythedifferentiatedBMSCswithneuronspecificenolase(NSE).ResultsAftersevendaysofculture,someBMSCsingrupeAappearedtypicalmorphologicalneuronaltraits.Cellbodiesappearedrefractile,cone-likeorsphericalwithlongprocesses,NSEwas
7、expressedingroupA.ThecellsingroupBandgroupCstillhadnoNSEexpression.ConclusionCombinationofbFGF,EGFandRAcaninduceBMSCstodifferentiateintoNSCsandneurons,andcanregulatethedifferentiationandpromotethematurationofneurons.ButBMP-7cannotinduceBMSCstodifferentiateintoNSCsandneuronsatthiscond
8、ition.��Keyw