自体血清与胎牛血清培养口腔黏膜角质细胞及上皮组织的实验研究刘立强李森恺范金财李养群刘元波王永前徐家杰曹蕊周传德【摘要】目的探讨自体血清培养人口腔黏膜角质细胞的可行性,为组织工程口腔黏膜用于临床提供理论和技术依据。方法应用自行制备的含10%、20%和30%自体血清的培养液及10%胎牛血清培养液,对人口腔黏膜角质细胞进行培养。对比观察不同浓度自体血清和胎牛血清培养的口腔黏膜角质细胞及上皮组织生长情况,绘制10%自体血清组及10%胎牛血清组细胞生长曲线及计算其倍增时间。并行抗2HLA免疫荧光检测培养上皮。结果各浓度自体血清组和10%胎牛血清组细胞生长及形态无明显差别,10%自体血清组细胞倍增时间为24.02±1.80h,与10%胎牛血清组20190±0179h比较,差异无统计学意义(P>0.05。各组细胞24h克隆形成率比较,差异均无统计学意义(P>0105;随自体血清浓度升高获得黏膜上皮面积增大,厚度增加,以20%自体血清组最为明显,与10%胎牛血清组培养黏膜上皮比较,差异有统计学意义(P<0.05。各组培养上皮经抗2HLA免疫荧光检测为阳性。结论制备的自体血清能完全替代胎牛血清对口腔黏膜角质细胞进行培养,且培养的口腔黏膜上皮组织分化优于胎牛血清。【关键词】自体血清胎牛血清口腔黏膜角质细胞口腔黏膜上皮组织培养中图分类号:R318Q813.11文献标识码:ATHEEXPERIMENTALSTUDYONCULTUREOFHUMANORALKERATINOCYTEANDEPITHELIUMUSINGAUTOLOGOUSSERUMANDFETALBOVINESERUMLIULiqiang,LISenkai,FANJincai,etal.PlasticSurgeryHospitalofCAMS,Beijing,100041,P.R.ChinaCorrespondingauthor:LIULiqiang,E2mail:llqqt@yahoo.com.cn【Abstract】ObjectiveToinvestigatethepossibilityofculturinghumanoralkeratinocyteusingautologousseruminordertoprovidetheoreticalandtechnicalfoundationforclinicalapplicationoftissueengineeringoralmucosaepithelium.MethodsThehumanoralkeratinocyteswereculturedbythemediumcontainingdifferentconcentrationsofautologousserum(10%,20%,30%andfetalbovineserum(10%,respectively.Thegrowthconditionsforthecellandthemucosaepitheliuminthegroupswereobserved,thecellgrowthcurvesweredrawn,andthepopulationdoublingtime(PDTwascounted.ResultsTheresultsshowedthatthehumanoralkeratinocytecouldproliferatewellinthemediumcontainingautologousserumorfetalbovineserum.Thedifferencesinthe242hourclonerateandPDTwerenotsignificant.Boththeareaandthethicknessoftheculturedoralepitheliumincreasedwiththeincreaseoftheautologousserumconcentration,andthedifferencebetweenautologousserumandfetalbovineserumwassignificant,especiallywiththemediumcontaining20%autologousserum(P<0105.Thehumannatureoftheculturedepitheliumwasdemonstratedbytheimmunofluorescentmouseanti2HLAantigen.ConclusionTheautologousserumcanreplacethefetalbovineserumtoculturetheoralkeratinocytewell,andtheculturedoralmucosaepitheliumcanbebetterdifferentiatedintheautologousserumthaninthefetalbovineserum.【Keywords】AutologousserumFetalbovineserumOralkeratinocyteOralepitheliumCultureFoundationitems:NationalNaturalScienceFoundationofChina(30070779;HealthMinistryFoundationofChina(20010409血清因含有类似于体内环境条件下的多种促细胞生长营养成分和细胞因子,成为细胞培养的常规添加成分。常用血清为动物来源血清,如鸡、猪、牛等,其中基金项目:国家自然科学基金资助项目(30070779;卫生部临床学科重点资助项目(20010409作者单位:中国医学科学院中国协和医科大学整形外科医院(北京,100041通讯作者:刘立强,主治医师,研究方向:组织工程,E2mail:llqqt@yahoo.com.cn胎牛血清因获取量大、污染轻而被广泛应用,但近年随着人畜共患病发病率的增加,以异种血清培养的组织向临床应用的安全性受到质疑[1],因此自体血清培养越来越受到重视[2]。我们的研究旨在探讨自体血清培养人口腔黏膜细胞及上皮组织的可行性。1材料与方法1.1组织取材与试剂术中剩余口腔黏膜...