应用EB病毒转化技术建立拉祜族和普米族永生细胞库.pdfVIP

中文摘要 中国有 56个民族和许多遗传隔离群体，长期保存各民族基因组的主 要方法是建立中国各民族永生细胞库。本研究从云南省勋海县和兰坪县 采集了50个拉枯族和84个普米族健康个体的血样本，采用EB病毒转化 B淋巴细胞，建立了127个类淋巴母细胞系，转化率达95%。同时，培养 基培养法和PCR法检测 10个细胞株的支原体，结果显示没有发生支原体 污染。这表明本研究己成功建立起 2个民族永生细胞库，其传代培养、 细胞冻存、细胞复苏和支原体污染防范等一整套过程是满足建库基本要 求的。细胞生长和形态学观察及染色体分析结果证实，其转化细胞的生 物学特征及遗传学特征是稳定的，为今后基因组多样性和现代人群历史 的研究提供了实验依据。另外，本研究还对一些影响转化的因素和可能 的机理进行了探讨。 关键词:永生细胞库 类淋巴母细胞系支原体EB病毒PCR 英文摘要 Establishmentoflymphoblastoidcelllines byEBV-transformationofBlymphocytesfromtwo Chineseminorities Postgraduate: HuangXiaoqin Thesisadvisor:ProfessorChuTiayou InstituteofMedicalBiology,ChineseAcademyofMedicalSciences Abstract Thereare56minoritiesandmanygeneticallyisolatedpopulationsm . China.Therpain methodforlong-termpreservationofthegenomicsourceso f these lationsistoestablishanimmortalizedcellpanel.Inthisresearch,the Blymphocytesfrom134individualsinLahuandPumiminoritieslivingin MenghaiCountyandLanpingCounty,YunnanProvince, transformedby Epstein-Barrvirusandaresourcepanelof127lymphoblastoidcelllines (LCLs),withatrans ationrateof95%,wasestablished.Furthermore,10 LCLswere testedformycoplasmalcontamination勿 polymerasechain reaction-basedmethodandcultureassay,showingaresultofnegative infestation.Theseresearchresultssuggestedthataresourcepanelof即stein- Barrvirusimmortalizedcellsrfomtwominoritypopulationswassuccessfully established,andthattheprocessespertainingtosubculturing,cryogenic preservation,culturerecoveryandmycoplasmalcontaminationpreventionwere satisfactorilyadapted.Themicroscopicevidencesofcellgrowthpatternsand cellularmorphology,andchromosomalstructuresconfirmedacellularand geneticstabilityin 10Epstein-BarrvirustransformedLCLs,providing experimentaldataforstudiesofgenomicdiversityandhistoryofmodern humanpopulations.Inaddition,thepossiblemechanismsandsomefactors involvedincelltransformationarediscussedintheresear