Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/3483
標題: 利用葡萄聚糖接枝聚乙烯亞胺載體對動物細胞轉染效率之探討
作者: 唐建翔
Hsiang, Tang Chien
關鍵字: 葡萄聚糖;聚乙烯亞胺;基因傳遞;細胞轉染
出版社: 化學工程學系
摘要: 
In this study, dextran of molecular weight 1500 was grafted onto branched polyethylenimine ( PEI ). The dextran-grafted PEI was mixed with plasmid pEGFP-C1 to form complexes to transfect CHO and 231 cells. It was found that the transfection efficiency was enhanced when PEI was prepared by the reaction using low weight ratios of dextran to PEI ( 0.25 and 0.5 ). When EMA-labeled plasmid was used to examine plasmid delivery, it was found that the intracellular amounts of the delivered plasmid within CHO cells became less for using dextran-grafted PEI. The cytotoxicity of the dextran-grafted PEI was slighty reduced when the DNA / polymer complexes were used to transfect CHO cells. As the titration experiments revealed, the buffering capacity of the dextran-grafted PEI was higher than that of the unmodified PEI, and increased as the grafting degree decreased. The increased buffering capacity might cause the enhanced transfection efficiency.

本研究利用分子量為1500之葡萄聚醣,對支鏈型聚乙烯亞胺進行接枝反應。接枝後產生的支鏈型聚乙烯亞胺載體與質體核酸pEGFP-C1形成複合物,並分別對中國大頰鼠卵巢細胞與人類胸線癌細胞進行轉染,觀察其蛋白質表現效率的差異。結果發現葡萄聚醣/支鏈型聚乙烯亞胺以低接枝比例(0.25/1與0.5/1,w/w)所合成之載體,和質體核酸形成複合物後,對於動物細胞的轉染效率,較未接枝的支鏈型聚乙烯亞胺載體為高。該載體與經過EMA標記之質體核酸形成複合物,對中國大頰鼠卵巢細胞中進行細胞傳遞的測試發現,經過接枝後的支鏈型聚乙烯亞胺載體,其傳遞效率並未獲得提升,甚至略低於未接枝的載體傳遞效率。利用中國大頰鼠卵巢細胞進行細胞毒性測試,發現經過接枝反應的載體與質體核酸形成複合物並對動物細胞進行轉染後,其毒性較未接枝的載體進行轉染時來的低。另外,利用酸鹼滴定對接枝後支鏈型聚乙烯亞胺載體進行酸鹼緩衝能力的測試中(由酸鹼值10滴定至4)發現,經過接枝後的載體高分子,其對於酸鹼緩衝的能力,較未接枝的支鏈型聚乙烯亞胺來的高。而且葡萄聚醣/支鏈型聚乙烯亞胺之接枝比例越低,其對於酸鹼緩衝的能力就越高。這個現象或許是低接枝比例的支鏈型聚乙烯亞胺,與質體核酸形成複合物後,對動物細胞之轉染效率提升的主要原因。
URI: http://hdl.handle.net/11455/3483
Appears in Collections:化學工程學系所

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