|
OALib Journal期刊
ISSN: 2333-9721
费用：99美元
|
|
|
|
骨髓间充质干细胞的肌源性诱导分化及转染vegf基因的表达
, PP. 290-294
Keywords: 基因表达,动物,转基因/免疫学,骨髓间充质干细胞,肌源性细胞,血管内皮生长因子
Abstract:
目的体外诱导兔骨髓间充质干细胞向肌源性细胞分化,并观察血管内皮生长因子adtrackcmv-hvegf165真核表达质粒转染骨髓间充质干细胞(mesenchymalstemcells,mscs)后的表达。方法将20只兔随机分为对照组和实验组,从骨髓液中分离mscs,对照组以低糖dmem培养,实验组加用5-氮胞苷(10mmol/l)诱导培养,第28天进行肌钙蛋白i免疫组化染色。另构建adtrackcmv-hvegf165真核表达质粒,通过脂质体转染对照组mscs,用northernblotting和westernblotting鉴定血管内皮生长因子(vegf)的表达,并用elisa法测定培养上清液中vegf的浓度。结果成功分离培养出兔mscs,实验组经5-氮胞苷诱导后mscs向肌源性细胞分化,肌钙蛋白i免疫组化染色阳性。成功构建adtrackcmv-hvegf165真核表达质粒并通过脂质体转染mscs,northernblotting示转染的mscsvegf165表达信号明显强于未转染细胞,westernblotting示转染vegf165基因的msc表达vegf,elisa法测定培养上清液中vegf的浓度在转染后第3天(1011pg/ml)和第5天(1027pg/ml)达高峰,此后逐渐下降,但至第13天(349pg/ml)仍显著高于空白对照(116pg/ml)和padtrackcmv组(125pg/ml),p<0.01。结论兔骨髓间充质干细胞可在体外向肌源性细胞分化,转染vegf基因可表达vegf,有望将干细胞移植和基因治疗结合以治疗缺血性心脏病。
| [1] | pittengermf,mackayam,becksc,etal.multilineagepotentialofadulthumanmesenchymalstemcells[j].science,1999,284(5411):143-7.
|
| [2] | caplanai.mesenchymalstemcellsandgenetherapy[j].clinorthop,2000,379(suppl):s67-70.
|
| [3] | perinec,dohmannhf,borojevicr,etal.transendocardial,autologousbonemarrowcelltransplantationforsevere,chronicischemicheartfailure[j].circulation,2003,107(18):2294-302.
|
| [4] | meinell,karageorgiouv,fajardor,etal.bonetissueengineeringusinghumanmesenchymalstemcells:effectsofscaffoldmaterialandmediumflow[j].annbiomedeng,2004,32(1):112-22.
|
| [5] | 林竞韧,郭坤元,李江琪,等.人骨髓基质于细胞克隆的培养及其向神经元样细胞的定向诱导分化[j].第一军医大学学报,2003,23(3):251-3.linjr,guoky,lijq,etal.invitrocultureofhumanbonemarrowmesenchymalstemcellcloniesandinduceddifferentiationintoneuron-likecells[j].jfirstmilmeduniv/diyijunyidaxuexuebao,2003,23(3):251-3.
|
| [6] | wakitanis,saitot,caplanai.myogeniccellsderivedfromratbonemarrowmesenchymalstemcellsexposedto5-azacytidine[j].musclenerve,1995,18(12):1417-26.
|
| [7] | brittenmb,abolmaalind,assmusb,etal.infarctremodelingafterintracoronaryprogenitorcelltreatmentinpatientswithacutemyocardialinfarction(topcare-ami):mechanisticinsightsfromserialcontrast-enhancedmagneticresonanceimaging[j].circulation,2003,108(18):2212-8.
|
| [8] | fuchss,satlerlf,komowskir,etal.catheter-basedautologousbonemarrowmyocardialinjectioninno-optionpatientswithadvancedcoronaryarterydisease:afeasibilitystudy[j].jamcollcardiol,2003,41(10):1721-4.
|
| [9] | 江逊,崔鹏程,陈文弦,等.诱导的人骨髓间充质干细胞在裸鼠体内生成软骨的实验研究[j].第一军医大学学报,2003,23(8):766-9.jiangx,cuipc,chenwx,etal.invivochondrogenesisofinducedhumanmarrowmesenchymalstemcellsinnudemice[j].jfirstmilmeduniv/diyijunyidaxuexuebao,2003,23(8):766-9.
|
| [10] | 路艳蒙,傅文玉,朴英杰.人骨髓间充质干细胞的培养及性质鉴定[j].第一军医大学学报,2001:21(8):571-3.luym,fuwy,piaoyj.cultureandidentificationofhumanmesenchymalstemcells[j].jfirstmilmeduniv/diyijunyidaxuexuebao,2001:21(8):571-3.
|
| [11] | makinos,fukudak,miyoshis,etal.cardiomyocytescanbegeneratedfrommarrowstromalcellsinvitro[j].jclininvest,1999,103(5):697-705.
|
| [12] | hassinkrj,bruteldelarivierea,mummerycl,etal.transplantationofcellsforcardiacrepair[j].jamcollcardiol,2003,41(5):711-7.
|
| [13] | tomitas,lirk,weiselrd,etal.autologoustransplantationofbonemarrowcellsimprovesdamagedheartfunction[j].circulation,1999,100(19suppl):ii247-56.
|
| [14] | 金丹,曾位森,裴国献,等.经转染人骨形态发生蛋白7基因后骨髓间充质干细胞mrna的表达[j].第一军医大学学报,2002,22(3):222-5jind,zengws,peigx,etal.expressionofhumanbonemorphogeneticprotein7mrnaafterthegenetransfectioninrabbitbonemarrow-derivedmesenchymalstemcells[j].jfirstmilmeduniv/diyijunyidaxuexuebao,2002,22(3):222-5.
|
| [15] | yautm,fungk,weiselrd,etal.enhancedmyocardialangiogenesisbygenetransferwithtransplantedcells[j].circulation,2001,104(12suppl1):i218-22.
|
Full-Text
|
|
Contact Us
service@oalib.com QQ:3279437679 
WhatsApp +8615387084133
|
|
