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研究生:陳莞婷
研究生(外文):Wan-Ting Chen
論文名稱:以醣類複合性幾丁聚醣顆粒培養動物細胞生產病毒
論文名稱(外文):Using sugar-composite chitosan particles for animal cell culture and virus production
指導教授:林雅菁林雅菁引用關係
指導教授(外文):Ya-Ching Lin
學位類別:碩士
校院名稱:輔英科技大學
系所名稱:生物技術系碩士班
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2008
畢業學年度:96
語文別:中文
論文頁數:75
中文關鍵詞:醣類幾丁聚醣細胞培養病毒培養
外文關鍵詞:ChitosanSugarVirus productionCell culture
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幾丁聚醣 (chitosan)是由幾丁質經去乙醯作用(deacetylation)反應後所得到的產物。幾丁聚醣具有許多特性例如:生物相容性、生物可分解性、抗菌性及易吸附細胞表面不會產生細胞毒性,已被廣泛應用於組織工程的基材。本實驗利用幾丁聚醣結合不同濃度10、20 以及30%的葡萄糖、蔗糖及澱粉形成醣類複合性幾丁聚醣顆粒,進行細胞吸附實驗。其中以含澱粉30%之醣類複合性幾丁聚醣顆粒對於非洲綠猴腎臟細胞 (Vero cell) 吸附性與細胞生長效果最好,而猪腎傳代細胞(PK15 cell)的吸附性與細胞生長效果最好為含澱粉10%之醣類複合性幾丁聚醣顆粒。利用這兩種顆粒分別進行腸病毒及猪瘟病毒培養,其腸病毒產量比一般培養皿為高,同時病毒亦容易被釋放於細胞外,未來可利用此基質於生物反應器進行病毒大量生產。
Chitosan is a natural nontoxic biopolymer derived by the deacetylation of chitin. Chitosan has properties of biocompatibility, biodegradability,antibacterial activity and ease to adsorb the surface of cells. With these
characteristics, it has attracted much attention as a new matrix for cell culture engineering. In this study, sugar-composite chitosan particles including glucose-chitosan particles, sucrose-chitosan particles and starchchitosan
particles with varied concentrations of sugar were prepared for the culture of Vero and PK15 cells. Our results showed that among these particles, 30% starch-chitosan particles were the best carriers for Vero cells
growth while 10% starch-chitosan particles were the best carriers for PK15 cells. Virus production in these two types of starch-chitosan particles, particular for enterovirus production, was better than that in culture flasks, suggesting promising application of the starch-chitosan particles to virus vaccine production.
誌謝 -----------------------------------------------------i
中文摘要 -----------------------------------------------iii
英文摘要 ------------------------------------------------iv
目錄 -----------------------------------------------------v
表目錄 ------------------------------------------------ vii
圖目錄 ----------------------------------------------- viii
第一章 緒論---------------------------------------------- 1
第二章 文獻探討-------------------------------------------2
第一節 幾丁質與幾丁聚醣之介紹-----------------------------2
第二節 載體上動物細胞培養於生物反應器---------------------4
第三節 醣類對細胞培養生長的影響-------------------------- 8
第四節 腸病毒71 型簡介------------------------------------9
第五節 猪瘟病毒簡介--------------------------------------10
第三章 材料與方法----------------------------------------12
第一節 實驗材料------------------------------------------12
第二節 醣類幾丁聚醣複合顆粒之製備------------------------13
第三節 細胞株之培養--------------------------------------13
第四節 病毒株之培養--------------------------------------14
第五節 病毒力價之測試 (TCID50)---------------------------14
第六節 葡萄糖測定----------------------------------------16
第七節 實驗流程規劃--------------------------------------16
第八節 統計學計算----------------------------------------20
第四章 結果與討論----------------------------------------21
第一節 不同醣類複合性幾丁聚醣顆粒之特性------------------21
第二節 不同醣類複合性幾丁聚醣顆粒上細胞貼附之情形--------21
第三節 不同醣類複合性幾丁聚醣顆粒上Vero 細胞生長情形---- 22
第四節 不同醣類複合性幾丁聚醣顆粒上PK15 細胞生長情形--- 25
第五節 醣類複合性幾丁聚醣顆粒應用於病毒的生產------------28
第六節 不同形狀的澱粉幾丁聚醣顆粒細胞生長之測試----------29
第七節 生物反應器的應用----------------------------------31
第五章 結論與建議----------------------------------------32
參考文獻 ------------------------------------------------34
附錄一 葡萄糖濃度校正曲線之標準溶液配置表----------------75
附錄二 葡萄糖測定的DNS 試劑配製表------------------------75
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