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利用超音波增強小鼠纖維母細胞重組為誘導型多潛能性幹細胞 = Ultras...

國立高雄大學生物科技研究所

利用超音波增強小鼠纖維母細胞重組為誘導型多潛能性幹細胞 = Ultrasound enhances mouse fibroblast Ultrasound enhances mouse fibroblast reprogramming to induced pluripotent stem cells

紀錄類型:	書目-語言資料,印刷品 : 單行本
並列題名:	Ultrasound enhances mouse fibroblast Ultrasound enhances mouse fibroblast reprogramming to induced pluripotent stem cells
作者:	廖專琪,
其他團體作者:	國立高雄大學
出版地:	[高雄市]
出版者:	撰者;
出版年:	2011[民100]
面頁冊數:	103面圖，表格 : 30公分;
標題:	磷脂體
標題:	liposomes
電子資源:	http://handle.ncl.edu.tw/11296/ndltd/59272033045459723846
附註:	105年10月25日公開
附註:	參考書目：面64-71
摘要註:	利用轉染載體將誘導型多潛能性幹細胞(iPS細胞)轉錄因子(Oct4、Sox2、Klf4、c-myc)送入體細胞後，體細胞會因基因重組成而轉變成另一種細胞，此種細胞特徵與胚胎幹細胞類似，具有相同的基因表現與生長特性，同樣擁有自我更新且分化成三胚層細胞的能力，因此被命名為iPS細胞。目前最常用來製備iPS細胞的基因轉染方式為慢病毒載體，轉染效率僅約0.1%~1%。超音波是另一種新的轉染方式，當超音波在液體中傳遞時，會因空穴現象(cavitation)而產生真空小泡，真空小泡破裂時所釋放出來的壓力，能有效的攻擊細胞膜，增加細胞通透性，藉此增進基因轉染效率。本研究先以不同粒徑磷脂體模擬細胞與反轉錄病毒，證實在超音波刺激下，能夠促進磷脂體通透性，並釋放內容物，以界面電位分析證實超音波並不會造成磷脂體化學組成的改變，利用calcein-磷脂體與GFP-慢病毒載體配合超音波作用於小鼠纖維母細胞，發現給予超音波刺激的組別，能促進calcein-磷脂體貼附在小鼠纖維母細胞上，並且GFP-慢病毒載體基因轉染率為23.81%，比未施加超音波的組別轉染率多了2.27倍，以iPSC–慢病毒載體感染小鼠纖維母細胞，發現有施加給予超音波的對照組比起未施加超音波的控制組，其iPS細胞聚落(colonies)數量增加了40%，觀察iPS細胞型態，並配合DNA電泳，免疫螢光染色法，拉曼圖譜分析，證實iPS細胞表現特有的細胞特徵，實驗結果發現超音波確實能增加慢病毒載體基因轉染率，進而增加iPS細胞聚落數，在未來可以利用超音波配合其他基因轉染方式，增強iPS細胞表現。 The somatic cell could reprogram to induced pluripotent stem cell (iPSC) by gene transfection vector. The characters of iPS cells are like embryonic stem cells (ESC), they have the same gene expression and life characteristic, include self-renew and differentiate to three germ layers. The preparation of iPSC is by lentiviral vector, but the transfect rate of is only 0.1 to 1%. Ultrasound is a new method of gene transfection. When ultrasound transmits energy in fluid, it could cause cavitations and damage the cell membrane. Therefore, ultrasound could enhance the cell permeability and then increase the gene transfecrion rate. In this study, different sizes of liposomes were used to mimic cells and lentivirus. The permeability of liposomes was increased during insonation. Same phenomena were found for applying sound waves on mouse fibroblast in presence of calcein-liposomes and GFP-lentiviral vectors. Sound waves could enhance the binding of calcein-liposomes and increase the transfection of GFP-lentiviral vectors to fibroblast. The transfection efficiency of GFP-lentivires is 23.81% which 2.27-folds higher than that without ultrasound. Same protocol was employed for the transfection of iPSC. The iPS cellular colonies number is 40% higher than without sound treatment. The expression of iPS cells is then identified by cellular morphology, DNA electrophoresis, immunofluroescence and Raman microscopy. The result showed that ultrasound could enhance lentiviral vector gene transfection and then increase iPS cells colonies.

利用超音波增強小鼠纖維母細胞重組為誘導型多潛能性幹細胞 = Ultrasound enhances mouse fibroblast Ultrasound enhances mouse fibroblast reprogramming to induced pluripotent stem cells
廖, 專琪

利用超音波增強小鼠纖維母細胞重組為誘導型多潛能性幹細胞 = Ultrasound enhances mouse fibroblast Ultrasound enhances mouse fibroblast reprogramming to induced pluripotent stem cells / 廖專琪撰 - [高雄市] : 撰者, 2011[民100]. - 103面 ; 圖，表格 ; 30公分.
105年10月25日公開參考書目：面64-71.
磷脂體liposomes

利用超音波增強小鼠纖維母細胞重組為誘導型多潛能性幹細胞 = Ultrasound enhances mouse fibroblast Ultrasound enhances mouse fibroblast reprogramming to induced pluripotent stem cells
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