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Title: | 以幾丁聚醣薄膜作為癌細胞與人類誘導多能幹細胞之新穎三維細胞培養平台 Chitosan Membranes as A Novel 3D Cell Culture Platform for Cancer Cells and Human Induced Pluripotent Stem Cells |
Authors: | Po-Hsiang Chang 張博翔 |
Advisor: | 陳彥榮(Edward Chern) |
Keyword: | 幾丁聚醣,三維細胞培養系統,癌症幹性,人類誘導多能幹細胞, chitosan,3D cell culture system,cancer stemness,human induced pluripotent stem cell, |
Publication Year : | 2020 |
Degree: | 博士 |
Abstract: | 二維細胞培養常作為細胞生物學與醫學領域的研究模式,但生物體內的各種組織與器官處於複雜的三維網絡中,因此相較於二維細胞培養,三維細胞培養平台能準確地模擬體內真實的狀態。過去研究指出透明質酸與幾丁聚醣複合膜可作為肺癌細胞、胰臟癌細胞與間葉幹細胞的簡易培養平台。本論文則開發以幾丁聚醣為主的三維細胞培養系統,用於培養大腸癌細胞、肝癌細胞與人類誘導多能幹細胞。本論文第一部份,幾丁聚醣薄膜會促進癌細胞的腫瘤進程與幹細胞特性。除此之外,幾丁聚醣可能會活化 CD44 陽性大腸癌中典型的 Wnt/β-catenin-CD44 訊息路徑,也活化 CD44 陰性肝癌中非典型的 Wnt-STAT3 的訊息路徑。第一部分的研究顯示,幾丁聚醣作為培養基材能促進癌細胞的幹細胞特性,以作為未來研究癌症的基礎。基於幾丁聚醣與細胞幹性的交互作用,此薄膜有潛力成為簡便且符合成本效益的誘導多能幹細胞的培養平台。本論文第二部份,人類誘導多能幹細胞於幾丁聚醣薄膜上能長期維持住其增生速率與多能性,甚至相比於作為控制組的玻連蛋白基質,幾丁聚醣能促進類似於原態多能性的表徵。人類誘導多能幹細胞會於幾丁聚醣薄膜上自組裝形成三維球體,這些球體於此薄膜上可直接誘導成具有三維結構的三胚層細胞。第二部分的研究顯示,幾丁聚醣薄膜不僅能促進人類誘導多能幹細胞的類原態多能性特徵,也能作為一個新穎的三維分化模式。總結來說,此簡便以生醫材料為底的系統能作為一個癌細胞與誘導多能幹細胞的簡便三維培養平台,希望能加速再生醫學、疾病模擬與藥物開發的未來發展。 Two-dimensional (2D) adherent cell culture is widely used for research model in cell biology and medicine. However, tissues and organs are constructed through cell-cell and cell-extracellular matrix connections with complex three-dimensional (3D) networks in vertebrates. Emerging evidence indicated that 3D cell culture systems could mimic in vivo conditions and provide the accurate biological properties of cells. Previous studies have demonstrated that chitosan membranes crosslinked with hyaluronic acid could be a simple non-adherent 3D cell culture platform to study lung cancer cells, pancreatic cancer cells, and mesenchymal stem cells. In this dissertation, chitosan-based 3D cell culture system would be developed for the culture of colon cancer cells, hepatocellular carcinoma (HCC) cells, and human induced pluripotent stem cells (hiPSCs). In the first part, chitosan membranes promoted tumor progression and stemness properties. Furthermore, chitosan could activate canonical Wnt/β-catenin-CD44 axis signaling in CD44positive colon cancer cells and noncanonical Wnt-STAT3 signaling in CD44negative HCC cells. Chitosan as a simple culture substrate can regulate cancer stemness for further cancer research and drug screening. On the other hand, based on the relation between chitosan and cell stemness, chitosan probably served as a simplified and cost-effective culture system for hiPSCs. In the second part, chitosan membranes sustained the proliferation and pluripotency of hiPSCs in long-term culture. Moreover, using vitronectin as the comparison group, hiPSCs grown on the membranes displayed naïve-like characteristics. On the chitosan membranes, hiPSCs self-assembled into 3D spheroids which could be directly differentiated into lineage-specific cells from the three germ layers with 3D structures. Chitosan membranes not only promoted the naïve pluripotent features of hiPSCs but also provided a novel 3D differentiation platform. Collectively, this convenient biomaterial-based culture system can provide a convenient platform to study cancer cells and hiPSCs and accelerate the development of regenerative medicine and disease modeling. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/74648 |
DOI: | 10.6342/NTU202100071 |
Fulltext Rights: | 有償授權 |
Appears in Collections: | 生化科技學系 |
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U0001-1601202112375100.pdf Restricted Access | 33.04 MB | Adobe PDF |
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