CAJ | 학술논문

体外构建PP2A Cα敲除的原代小鼠心室肌细胞模型。选择性地将雄性PP2A Cαfl/fl,Cre/-小鼠与雌性PP2A Cαfl/fl,Cre/-小鼠交配。新生小鼠心脏经胰酶消化后差速贴壁获得原代小鼠心室肌细胞,用带有Cre的腺病毒侵染心肌细胞,经荧光显微镜观察和Western-blot检测,分析PP2A Cα的敲除效果。将基因型为PP2A Cαfl/fl,Cre/-的雌雄小鼠进行交配,得到其同样基因型的后代,进而可获得足量基因型为PP2A Cαfl/fl,Cre/-的原代小鼠心肌细胞。用带有重组酶Cre的腺病毒侵染细胞48 h后,可见带有绿色荧光的心肌细胞；Western-blot检测显示,PP2A Cα在Cre腺病毒侵染的心肌细胞可下调60%~80%。本研究成功建立了PP2A Cα敲除的原代小鼠心肌细胞模型。
체외구건PP2A Cα고제적원대소서심실기세포모형。선택성지장웅성PP2A Cαfl/fl,Cre/-소서여자성PP2A Cαfl/fl,Cre/-소서교배。신생소서심장경이매소화후차속첩벽획득원대소서심실기세포,용대유Cre적선병독침염심기세포,경형광현미경관찰화Western-blot검측,분석PP2A Cα적고제효과。장기인형위PP2A Cαfl/fl,Cre/-적자웅소서진행교배,득도기동양기인형적후대,진이가획득족량기인형위PP2A Cαfl/fl,Cre/-적원대소서심기세포。용대유중조매Cre적선병독침염세포48 h후,가견대유록색형광적심기세포；Western-blot검측현시,PP2A Cα재Cre선병독침염적심기세포가하조60%~80%。본연구성공건립료PP2A Cα고제적원대소서심기세포모형。
To generate PP2A Cαknockout model in neonatal mouse ventricular myocytes. Male PP2A Cαfl/fl,Cre/-were se-lectively mated with female PP2A Cαfl/fl,Cre/- mice to obtain abundant offsprings with the same genotype. The neonatal mouse ventricular myocytes were digested by trypsinization and subsequently infected with Cre-adenovirus. The effects of adenovirus infection were observed by fluorescence microscope and identified by western blot analysis. The offsprings in aboundance were obtained by seting up a mating between male and female of PP2A Cαfl/fl,Cre/-mice, as a result, an abundance of neonatal mouse ventricular myocytes with the genotype of PP2A Cαfl/fl,Cre/-were available for Ad-Cre infection. Green fluorescence was observed after GFP labeled Cre-adenovirus( Cre-Ad) infection for 48 h. The expression level of PP2A Cα protein in Cre-Ad infected myocytes was 60% ~80% lower than control. The PP2A Cα knockout model in neonatal mouse ventricular myocytes was successfully generated.