CAJ | 학술논문

目的：建立新生SD大鼠c-kit阳性细胞分离纯化及传代的培养方法。方法：取出生24h之内新生SD大鼠心尖部组织，用PBS液洗去心尖部的血液，将其剪碎成lmm3以下的组织块培养，培养48h后，0.5％Trypsin-EDTA消化，用免疫磁珠分选c-kit阳性的心肌细胞，继续培养传代。用倒置显微镜观察c-kit阳性心肌细胞，用免疫细胞化学法检测心肌组织特异性cTnT及Desmin蛋白。结果：用心脏组织培养法成功得到了体外搏动的原代心肌细胞；用免疫磁珠分选了c-kit阳性心肌细胞，倒置显微镜下观察到c-kit阳性心肌细胞具有传代增殖的特性；同时，检测了c-kit阳性心肌细胞胞质cTnT及Desmin结合蛋白的阳性表达。结论：采用改良后的培养方法获得了c-kit阳性心肌细胞，该细胞具有心肌干细胞的蛋白表达特性，并具有一定的传代增殖能力。
목적：건립신생SD대서c-kit양성세포분리순화급전대적배양방법。방법：취출생24h지내신생SD대서심첨부조직，용PBS액세거심첨부적혈액，장기전쇄성lmm3이하적조직괴배양，배양48h후，0.5％Trypsin-EDTA소화，용면역자주분선c-kit양성적심기세포，계속배양전대。용도치현미경관찰c-kit양성심기세포，용면역세포화학법검측심기조직특이성cTnT급Desmin단백。결과：용심장조직배양법성공득도료체외박동적원대심기세포；용면역자주분선료c-kit양성심기세포，도치현미경하관찰도c-kit양성심기세포구유전대증식적특성；동시，검측료c-kit양성심기세포포질cTnT급Desmin결합단백적양성표체。결론：채용개량후적배양방법획득료c-kit양성심기세포，해세포구유심기간세포적단백표체특성，병구유일정적전대증식능력。
objective To establish a method about separation,purification and passage of c-kit+myocardial cells in SD newborn rats.Method Cardiac apex that removed from newborn SD rat within 24 hours washed by PBS,and cut it about lmm3,cultivate after 48 hours,0.5% Trypsin-EDTA digestion,c-kit+myocardial cells separated by positive immune magnetic bead ,continuing sub-culture.To observe the c -kit+myocardial cell growth,and immunocytochemistry detection of cTnT and Desmin protein.Results Pulsatile primary cardiomyocytes in vitro have been successfully gotten with cardiac tissue culture method;c -kit+myocardial cells have separated by positive immune magnetic bead,The passaged proliferation characteristics of c-kit-positive cardiac cells have been detected in an inverted microscope.Meanwhile,the positive expression of cTnT and Desmin in the cytoplasmic of c -kit-positive cardiac cells have been detected.Conclusion C-kit+myocardial cells,with the protein expression characteristics of myocardial stem cell and the proliferation ability ,have been obtained by the cultivation of the improved method.