CAJ | 학술논문

目的：探索纳米材料聚乙烯亚胺（ PEI ）携载不同分子量的蛋白质转染进人骨髓间充质干细胞（hBMSCs）的作用及优化其最佳配比条件。方法：利用物理化学作用构建6组PEI－DNase I（DNA酶I）复合体，运用激光散射技术初步摸索其最佳摩尔比，用透射电镜直观复原纳米－蛋白复合体外貌；同时，原代分离培养健康人的骨髓间充质干细胞并进行体外扩增，通过构建成功的4组PEI－GFP（绿色荧光蛋白）纳米蛋白复合体对hBMSCs进行转染，共聚焦荧光显微镜下观察荧光表达情况，再通过MTT法检测该复合体对细胞增殖的毒性影响，并用β－半乳糖苷酶实验验证其转入蛋白的活性表现。结果：当PEI与蛋白质的摩尔比为4∶1时，形成的复合体转染效率最高，β－半乳糖苷酶实验细胞染色变蓝。结论：PEI能与蛋白质形成纳米复合体，并能转染多种蛋白质进入人骨髓间充质干细胞，所转染的蛋白质分子仍具有活性，为细胞重编程技术提供了新途径。
목적：탐색납미재료취을희아알（ PEI ）휴재불동분자량적단백질전염진인골수간충질간세포（hBMSCs）적작용급우화기최가배비조건。방법：이용물이화학작용구건6조PEI－DNase I（DNA매I）복합체，운용격광산사기술초보모색기최가마이비，용투사전경직관복원납미－단백복합체외모；동시，원대분리배양건강인적골수간충질간세포병진행체외확증，통과구건성공적4조PEI－GFP（록색형광단백）납미단백복합체대hBMSCs진행전염，공취초형광현미경하관찰형광표체정황，재통과MTT법검측해복합체대세포증식적독성영향，병용β－반유당감매실험험증기전입단백적활성표현。결과：당PEI여단백질적마이비위4∶1시，형성적복합체전염효솔최고，β－반유당감매실험세포염색변람。결론：PEI능여단백질형성납미복합체，병능전염다충단백질진입인골수간충질간세포，소전염적단백질분자잉구유활성，위세포중편정기술제공료신도경。
[ ABSTRACT] AIM:To investigate the role of encapsulated protein transfected into human bone marrow mesen-chymal stem cells ( hBMSCs) by polyethyleneimine ( PEI) , and to optimize the best mole ratio of PEI-proteins.METH-ODS:6 groups of DNase I-PEI complexes were constructed and the best mole ratio was explored by laser scattering analy-sis.The appearance of complexes was presented under transmission electron microscope.Meanwhile, 4 groups of construc-ted GFP-PEI complexes were utilized to transfect into the hBMSCs, which were isolated and expand in vitro.The fluores-cence intensity of transfected cells was observed under confocal microscope.In addition, the cytotoxicity of the complexes on the cell proliferation was detected by MTT assay.The activity of the intracellular proteins was testified by aβ-galactosi-dase staining experiment.RESULTS:When the mole ratio of PEI and protein was adjusted to 4∶1, the complex transfec-tion efficiency was the best, and β-galactosidase color test turned blue.CONCLUSION:PEI has the character of encap-sulating various proteins to nano-complexes.The proteins transfected into bone marrow mesenchymal stem cells are con-firmed to have functional activity.As a protein carrier, PEI is of high efficiency and low toxicity, thus providing a new way for stem cell reprogramming.