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1、·1152·ChineseJournalofReparativeandReconstructiveSurgery,September2014,Vol.28,No.9白血病抑制因子聯(lián)合bFGF對(duì)人BMSCs增殖及分化的影響1，2，313132，32，32，3施劍明曹俊殷明殷嫦嫦程細(xì)高謝榮輝林思文徐浚懷【摘?要】目的?研究白血病抑制因子（leukemiainhibitoryfactor，LIF）聯(lián)合bFGF對(duì)人BMSCs（humanBMSCs，hBMSCs）增殖及分化的影響。?方法?取第4代hBMSCs，根據(jù)培養(yǎng)條件不同分為4組：A組為對(duì)照組，加入完全培養(yǎng)基（含10%FBS的α-MEM）常規(guī)培
2、養(yǎng)；B組加入含10ng/mLLIF的完全培養(yǎng)基；C組加入含10ng/mLbFGF的完全培養(yǎng)基；D組加入含10ng/mLLIF和10ng/mLbFGF的完全培養(yǎng)基。采用細(xì)胞計(jì)數(shù)試劑盒8（cellcountingkit8，CCK-8）測(cè)定各組第4代hBMSCs生長(zhǎng)曲線；倒置相差顯微鏡連續(xù)觀察各組hBMSCs形態(tài)變化；流式細(xì)胞術(shù)檢測(cè)各組第8代hBMSCs表面標(biāo)志CD44、CD90、CD19、CD34情況。?結(jié)果4組細(xì)胞生長(zhǎng)曲線均近似S形，且細(xì)胞增殖速率表現(xiàn)為D組>C組>B組>A組。連續(xù)觀察各組細(xì)胞形態(tài)發(fā)現(xiàn)A組細(xì)胞較早出現(xiàn)衰老和分化現(xiàn)象；B組細(xì)胞早期細(xì)胞形態(tài)維持較好，但細(xì)胞增殖相對(duì)緩慢，后期出現(xiàn)部
3、分衰老細(xì)胞；C組少量細(xì)胞呈神經(jīng)樣細(xì)胞分化，但增殖較快；D組細(xì)胞增殖迅速，且能長(zhǎng)期維持hBMSCs形態(tài)特征。流式細(xì)胞術(shù)檢測(cè)各組第8代hBMSCs表面標(biāo)志示：A、C組骨髓基質(zhì)標(biāo)志CD44、CD90表達(dá)較B、D組低；各組造血細(xì)胞標(biāo)志CD19、CD34均呈陰性，組間差異不明顯。結(jié)論LIF聯(lián)合bFGF不僅可以維持hBMSCs的多向分化潛能，而且能夠促進(jìn)其快速增殖?！娟P(guān)鍵詞】??白血病抑制因子??bFGF??人BMSCs??細(xì)胞增殖??細(xì)胞分化EFFECTSOFLEUKEMIAINHIBITORYFACTORCOMBINEDWITHBASICFIBROBLASTGROWTHFACTORONPROLIF
4、ERATIONANDDIFERENTIATIONOFHUMANBONEMARROWMESENCHYMALSTEMCELLS/SHI1,2,313132,32,32,31Jianming,CAOJun,YINMing,YINChangchang,CHENGXigao,XIERonghui,LINSiwen,XUJunhuai.BasicMedical23College,JiujiangUniversity,JiujiangJiangxi,332000,P.R.China;MedicineGraduateSchool,NanchangUniversity;DepartmentofOrthope
5、dics,theSecondAffiliatedHospitalofNanchangUniversity.Correspondingauthor:CAOJun,E-mail:caojun002@aliyun.com【Abstract】ObjectiveTostudytheeffectsofleukemiainhibitoryfactor(LIF)andbasicfibroblastgrowthfactor(bFGF)onproliferationanddifferentiationofhumanbonemarrowmesenchymalstemcells(hBMSCs).MethodshB
6、MSCsatpassage4weredividedinto4groupsaccordingtodifferentcultureconditions:cellsweretreatedwithcompletemedium(α-MEMcontaining10%FBS,groupA),withcompletemediumcontaining10ng/mLLIF(groupB),withcompletemediumcontaining10ng/mLbFGF(groupC),andwithcompletemediumcontaining10ng/mLLIFand10ng/mLbFGF(groupD).
7、ThegrowthcurvesofhBMSCsatpassage4indifferentgroupswereassayedbycellcountingkit8(CCK-8);cellularmorphologicchangeswereobservedunderinvertedphasecontrastmicroscope;thesurfacemarkersofhBMSCsatpassage8includingCD44,C
