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1、骨髓間充質(zhì)干細胞接種小鼠肝癌組織對小鼠生存期的影響作者:呂昕蕾,張南征,陳復(fù)興,劉軍權(quán),孫蕾清【摘要】目的研究骨髓間充質(zhì)干細胞(MSC)接種于原發(fā)性肝癌Heps荷瘤小鼠對其生存期的影響。方法體外貼壁培養(yǎng)法制備昆明小鼠骨髓MSC,分離培養(yǎng)傳代擴增,流式細胞儀鑒定MSC表型。4′,6-二脒基-2-苯基吲哚(4′,6-diamidino-2-phenylindole,DAPI)對MSC進行標(biāo)記備用。隨機取54只8周齡昆明小鼠,建立Heps肝癌移植瘤模型。移植瘤長徑達0.5~0.8cm時,隨機分為MSC組、DAPI標(biāo)記MSC組(DAPI組)、生理鹽水(NS)對照組(NS
2、對照組),每組18只。MSC組及DAPI組分別在小鼠Heps肝癌移植瘤內(nèi)注射2×106MSC及DAPI標(biāo)記的MSC,NS對照組注射同體積的NS。每組各有6只荷瘤小鼠用于觀察生存期。①自注射日起,隔日測量各組小鼠腫瘤長、短徑,計算注射后3、7、14及28天的腫瘤體積;②記錄注射當(dāng)天開始至小鼠死亡的時間,計算各組小鼠平均生存時間;③于注射后3、7、14及28天,各組分批處死3只小鼠。取腫瘤組織,石蠟包埋并切片,蘇木精-伊紅染色后光學(xué)顯微鏡觀察。結(jié)果①體外貼壁法成功制備MSC,流式細胞儀對第3代MSC進行鑒定,高表達CD29(97.84%),低表達CD34(5.56%
3、)、CD45(7.66%)。②小鼠Heps肝癌腹水瘤細胞成功制備小鼠Heps肝癌移植瘤模型,成瘤率100%。③注射MSC后1周,MSC組及DAPI組腫瘤體積增大明顯,與NS對照組差異有顯著性(P<0.05)。④MSC組平均生存期為45天(95%可信區(qū)間:33~56天),DAPI組平均生存期為34天(95%可信區(qū)間:31~37天),NS對照組平均生存期為33天(95%可信區(qū)間:28~37天);MSC組與NS對照組差異有顯著性(P<0.05),其余組間生存期差異無顯著性(P>0.05)。⑤各組小鼠腫瘤蘇木精-伊紅染色:28天時,MSC組及DAPI組
4、腫瘤組織壞死明顯,范圍較NS對照組更大。結(jié)論MSC可在原發(fā)性肝癌Heps荷瘤小鼠腫瘤組織內(nèi)定植,MSC接種后1周內(nèi)腫瘤增長迅速,小鼠生存期延長?!娟P(guān)鍵詞】肝腫瘤,實驗性;間充質(zhì)干細胞;Heps肝癌;小鼠Abstract:ObjectiveToinvestigatetheimpactofmesenchymalstemcells(MSC)inoculatedintotheestablishedmodelofHeps-bearingmiceonsurvivaltime.MethodsMSCethod.PhenotypesofMSCetry.MSCiceas.SCgro
5、up,MSClabeledalsaline(NS)controlgroup(NScontrolgroup).MSCandMSCmarkedbyDAPIinistratedintothemouserightrearbacktumortissueof2×106.TheNScontrolgroupinistratedevolumeofNS.6miceineachgroupe.①Thesizeoftheimplantedtumorseasuredevery2daysandthetumorvolumeeofthetumor-bearingmiceeansurvivalti
6、meicefromeachgrouportissuesamplescollectedatabove4timepointsbeddedinparaffinforroutinepathologicalsection,hematoxylin-eosinstainingandlightmicroscopyobservation.Results①PurifiedMSCethod.Passage3cellsetry.TheresultsshoodelofHeps-bearingmiceorformationis100%.③Atthefirstinistrated,thevo
7、lumeofthetumorsarkedlyincreasedintheMSCgroupandtheDAPIgroupparedeansurvivaltimeeansurvivaltimeeansurvivaltimeinistrated,thereortransplantationofHeps-bearingmice.InthefirsteoftumorintheMSCgroupisincreasedquickly.ThesurvivaltimeofHeps-bearingmiceisprolonged.Keys,experimental;mesenchyma
8、lstemcells;H