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1、小鼠骨髓間充質(zhì)干細(xì)胞多潛能性的鑒定第32卷第2期2011年4月暨南大學(xué)學(xué)報(醫(yī)學(xué)版)JournalofJinanUniversity(MedicineEdition)Vol.32Apr.No.22011黃11,21,2111偉,謝琪璇,秦俊文,李威娜,張春雪,肖鑾娟(暨南大學(xué)1.生命科學(xué)技術(shù)學(xué)院生殖免疫研宄所;2.再生醫(yī)學(xué)教育部重點實驗室,廣東廣州510632)[摘要]冃的:鑒定小鼠骨髓間充質(zhì)干細(xì)胞(MSCs)的多分化潛能的起因。方法:從成年小鼠骨髓股骨、脛骨中2、4、6、8、10d提取總RNA,PCR檢測多潛能特征基因和相
2、關(guān)因子、并進(jìn)行原代培養(yǎng),分別于0、通過RT-分離MSCs,3個胚層特征基因的mRNA的表達(dá)情況,4和以判斷MSCs的特性。結(jié)果:小鼠MSCs中表達(dá)多能性標(biāo)記基因Oct-nanog,Myc。夕卜胚層的nestin、并且表達(dá)與多能性相關(guān)的轉(zhuǎn)錄因子Klf4和c-中胚層的SM22a和內(nèi)胚層的CYP51標(biāo)記基因均有表達(dá)。結(jié)論:貼壁純化的小鼠MSCs除了具有多能性的干細(xì)胞,還可能含有各胚層的原始細(xì)胞。[關(guān)鍵詞]骨髓間充質(zhì)干細(xì)胞;[中圖分類號]Q786原代細(xì)胞培養(yǎng);RT-PCR;基因表達(dá)[文獻(xiàn)標(biāo)志碼]A[文章編號]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[基金項目]國家自然科學(xué)基金項目(30370541);211工程3期生物科學(xué)技術(shù)與生物醫(yī)學(xué)工程建設(shè)項目[作者簡介]黃偉(1984—),男,碩士研究生,研宄方向:生殖免疫與基因工程Tel:020—85220468;E-mail:txqx@jnu.edu.cn通