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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/16876
標題: | 利用第一型膠原蛋白凝膠與其培養出的三維嵌入球型人類結腸直腸癌細胞作為三維體外仿生模型之建立與評估 The Development of a Biomimetic In Vitro Model with 3D Spheres of Human Colorectal Cancer Cells embedded within Type I Collagen Gel |
作者: | Susan Yun Fan Lin 林芸帆 |
指導教授: | 謝銘鈞 |
關鍵字: | 仿生,二維/三維體外癌細胞模型,三維球型癌細胞,凝膠/支架嵌入,人類結腸直腸癌,第一型膠原蛋白凝膠,癌症幹細胞,幹性,上皮 - 間質轉化,血管新生,抗癌藥性,體內的致瘤性, biomimetic,2D/3D in vitro cancer cell model,3D cancer cell sphere,gel/scaffold-embedding,human colorectal cancer,type I collagen gel,cancer stem cell,stemness,epithelial-mesenchymal transition,angiogenesis,anti-cancer drug resistance,in vivo tumorigenicity, |
出版年 : | 2014 |
學位: | 碩士 |
摘要: | 體內和體外培養的癌細胞在許多表現上有著明顯的不同。為了更加了解進而治療癌症,這樣的落差必須被縮小。大部分的體外癌細胞模型是建立在二維的環境下,但是卻與體內的癌細胞之表現相去甚遠。而建立在三維環境下的體外癌細胞模型已被證實與體內的癌細胞有相似的表現。然而,卻沒有研究去探討所謂的三維環境下的體外癌細胞模型是否只需要具備三維的球型癌細胞或是還需將其培養在三維的微環境中。本研究使用人類結腸直腸癌細胞建立了三種體外癌細胞模型。分別為TCPS所培養的二維單層貼附癌細胞、超低附著表面所培養的三維懸浮非貼附球型癌細胞和由第一型膠原蛋白凝膠所培養的三維嵌入球型癌細胞。接著,這三種體外大腸癌細胞模型就各種體內癌細胞表現進行分析和比較並觀察哪一種模型更為接近體內癌細胞。實驗結果証實,由第一型膠原蛋白凝膠所培養出的三維嵌入球型癌細胞,其形態、活性、癌症幹細胞數量、幹性、上皮-間質轉化能力、血管新生、抗癌藥性和致瘤性都比其他兩種癌細胞模型還高。其次為由超低附著表面所培養的三維懸浮非貼附球型癌細胞,最後則是TCPS所培養的二維單層貼附癌細胞。由此可證,更為仿生的三維體外癌細胞模型不僅需要具備三維球型癌細胞也必須提供一個可讓三維球型癌細胞生長的三維微環境。 To better understand cancer, the gap between in vivo cancer cells and in vitro cancer cells must be narrowed. Such gap could be bridged by an in vitro cancer cell model that is biomimetic to in vivo cancer cells. While most in vitro cancer cell models are in two dimensions (2D), these 2D models are; however, also largely deviated from in vivo cancer cells. In vitro cancer cell models in three dimensions (3D) are proved to better recapitulate in vivo cancer cells. However, there is no study, so far, that specifies the prerequisites of a biomimetic 3D in vitro cancer cell model. Thus, this study aims to define whether a biomimetic 3D in vitro cancer cell model simply requires 3D cancer cell sphere growth or it also demands the 3D cancer cell sphere growth to be contained within a microenvironment. In this study, three in vitro cancer cell models were developed using human colorectal adenocarcinoma cells. A 2D monolayer model was constructed to comprise adherent 2D cancer cell monolayers cultured on tissue culture polystyrene (TCPS). A 3D spheroid model was created to consist suspension of non-adherent 3D cancer cell spheres cultured on ultra-low attachment surface (ULAS). A 3D gel/scaffold-embedding model was established to contain 3D cancer cell spheres cultured within the type I collagen gel (COL I) specially formulated in this study. Phenotypes of in vivo cancer cells including morphology, viability, cancer stem cell population, stemness, epithelial-mesenchymal transition (EMT), angiogenesis, anti-cancer drug resistance, and in vivo tumorigenicity were analyzed and compared among the three in vitro colorectal cancer cell models developed. The cell morphology of in vivo cancer cells was best duplicated by the 3D cancer cell spheres within COL I followed by the suspension of non-adherent 3D cancer cell spheres on ULAS and then the adherent 2D cancer cell monolayers on TCPS. The cell viability, cancer stem cell population, stemness markers expression, EMT markers expression, angiogenesis marker expression, anti-cancer drug resistance, and in vivo tumorigenicity of the 3D cancer cell spheres within COL I were also the highest followed by the suspension of non-adherent 3D cancer cell spheres on ULAS and then the adherent 2D cancer cell monolayers on TCPS. These results indicated that the 3D cancer cell spheres within COL I were the most biomimetic to in vitro cancer cells followed by the suspension of non-adherent 3D cancer cell spheres on ULAS and then by the adherent 2D cancer cell monolayers on TCPS. Therefore, it could be concluded that for the development of a more biomimetic 3D in vitro cancer cell model, not only 3D cancer cell sphere growth is needed but the 3D cancer cell sphere growth within a microenvironment is also necessitated. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/16876 |
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顯示於系所單位: | 醫學工程學研究所 |
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