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1、萬方數(shù)據(jù)Chin_,MarDrugs,2005-,“聊。V01.24No.3中國海洋藥物雜志2005年6月第24卷第3期海藻糖對低溫保存的氣管組織細胞活力的影響齊戰(zhàn)”,王勇杰1。王善政1,何奇1,邵軍2,呂麗紅2,尹金嶺2(1.山東大學齊魯醫(yī)院胸外科,山東濟南250012;2.山東大學醫(yī)學院手術學研究室,山東濟南250012)摘要:目的探討海藻糖對低溫保存的氣管組織細胞活力的影響。方法新鮮配制兩組保存波,其中:I組LPD+DMS0,Ⅱ組LPD+DMS0+海藻糖(O.10rn01.L_1)。切取SD大鼠氣管后立即分別放入含上述兩
2、種溶液的凍存管,在程序降溫儀降至一80℃后投入波氮中保存,分別在1,15。30,60,120d后對氣管組織體外培養(yǎng),并加入3H—TdR做標記,檢測細胞摻入率。結果采用3H-TdR體外組織培養(yǎng),低溫保存后I組氣管3H—TdR摻入率(88.72%~78.21%)明顯高于Ⅱ組(80.99%~70.75%),這種趨勢可長時間持續(xù)(共120d)。結論含有海藻糖的LPD溶液在低溫下對氣管組織細胞有明顯的保護作用;3H-TdR體外組織培養(yǎng)的方法可以比較客觀地反映了器官保存后的活力。.關鍵詞:海藻糖;氣管低溫保存;氣管組織細胞活力;3H—Td
3、R中圖分類號:R931.711,R96文獻標識碼:A文章編號:1002.3461(2005)03-0022.04EffectsoftrehaloseontrachealtissueandcellviabilitiesaftercryopreserVationQIZhanl,WANGYong—jiel,WANGShan-zhen91,HEQil,SHAOJun2,LULi—hon92,YINJin-lin92(1.DepartmentofThoracicSurgery,QiluHospital,ShandongUniversit
4、y,Jinan250012,China;2.DepartmentofOperationResearch,MedicalCollege,ShandongUniversity,Jinan250012,Chi—na)Abstract:ObjectiveTodetecttheeffectsoftrehaloseontrachealtissueandcellviabilitiesaf-tercryopreservation.MethodsInbredmaleSprague-Dawley(SD)ratsweresacrificedwith
5、intraperitonealinjectionofketamine(150mg·kg一1).Thetracheaswereremovedandim—mersedimmediatelyinthefreezingmediumoflOWpotassiumdextran(LPD)solutioncontai—ningwithlO%dimethylsulfoxide(DMSO)(GroupI)andcontainingwith10%DMSOandO.10molL.1trehalose(Group1I)respectively.Aste
6、rileplastictubecontaininga卜em—longtracheawasfilledwiththefreezingmedium,sealed,andfrozento一80℃atrateof一1℃perminuteinaprogrammablefreezer.Thenthetubewasstoredinliquidnitrogen(一196℃)fordifferentperiodsofpreservation(1,15,30,60,120d,n一---5).Forviabilityassessment,thesp
7、ecimenwasthawedat37℃inanincubatorandrinsedwithphysiologicsalinesolution10times.Next,eachspecimenwastrimmedforequalweight(100mg)andincubatedin3mLofRPMll640mediumcontaining10%fetalbovineserumat37℃for24hours.Thenthetra-cheaswerelabeledwith39Ci3H—TdRfor16hours.Thespecim
8、enswerehydrolyzedin80%formicacid(0.1mL)andincubatedati00℃for2hours.Atlast,thesolutionwascountedbyliquidscintillationcounter.ResultsTheaver