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1、小鼠骨髓间充质干细胞多潜能性的鉴定第32卷第2期2011年4月暨南大学学报(医学版)JournalofJinanUniversity(MedicineEdition)Vol.32Apr.No.22011黄11,21,2111伟,谢琪璇,秦俊文,李威娜,张春雪,肖銮娟(暨南大学1.生命科学技术学院生殖免疫研宄所;2.再生医学教育部重点实验室,广东广州510632)[摘要]冃的:鉴定小鼠骨髓间充质干细胞(MSCs)的多分化潜能的起因。方法:从成年小鼠骨髓股骨、胫骨中2、4、6、8、10d提取总RNA,PCR检测多潜能特征基因和相
2、关因子、并进行原代培养,分別于0、通过RT-分离MSCs,3个胚层特征基因的mRNA的表达情况,4和以判断MSCs的特性。结果:小鼠MSCs中表达多能性标记基因Oct-nanog,Myc。夕卜胚层的nestin、并且表达与多能性相关的转录因子Klf4和c-中胚层的SM22a和内胚层的CYP51标记基因均有表达。结论:贴壁纯化的小鼠MSCs除了具有多能性的干细胞,还可能含有各胚层的原始细胞。[关键词]骨髓间充质干细胞;[中图分类号]Q786原代细胞培养;RT-PCR;基因表达[文献标志码]A[文章编号]1000-9965(20
3、11)02-0136-05Theidentificationofmultipotentialinmousebonemarrow-derivedmesenchymalstemcells22HUANGWeil,XIEQi-xuanl,,QINJun-wenl,,LlWei-nal,ZHANGChun-xuel,XIAOLuan-juanl(1.InstituteofReproductivelmmunology,CollegeofLifeScienceandTechnology;2.KeyLaboratoryJinanUniver
4、sity,Guangzhou510632,China)forRegenerativeMedicineofMinistryofEducation,[Abstract]Aim:Toidentifythecauseofmulti-differentiationpotentialinmousebonemarrowmesen-chymalstemcells.Methods:MSCswereisolatedfromthefemursandtibiaeoftheadultmouseandcul-turedinvitro.TotaIRNAw
5、ereextractedfromthemousebonemarrow(DayO)andculturedmouseMSCsat2,4,6,8andl0days.ThepluripotencymarkergenesOct-4andnanog,pluripotencyrelatedtranscrip-tionfactorsKlf4andc-Mycandthemarkergenesofthreeembryonallayers,suchasCYP51,SM22aandnestin,whicharetypicalgenesspecifi
6、cforendoderm,mesodermandectodermrespectively,weredetectedbyRT-PCRinordertodeterminethecharacteristicsofMSCs.Results:TheresultsofRT-PCRshowedthatmouseMSCscanexpressmRNAofOct-4,nanog,Klf4andc-Myc.Furthermore,inadditiontoSM22a,RT-PCRrevealedthepresenceofmRNAforCYP51an
7、dnestinduringtheprimarycultureofmouseMSCs.Conclusion:MouseMSCs,whichwereisolatedbyadhesionpurification,maybeconsistofpluripo-tentsterncellsandprimarycellsfromthethreegermlayers.[Keywords]mesenchymalstemcells;expressionPrimarycellculture;ReversetranscriptionPCR;Gene
8、[收稿日期]2010-10-22[基金项目]国家自然科学基金项目(30370541);211工程3期生物科学技术与生物医学工程建设项目[作者简介]黄伟(1984—),男,硕士研究生,研宄方向:生殖免疫与基因工程Tel:020—85220468;E-mail:txqx@jnu.edu.cn通