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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 謝銘鈞 | |
dc.contributor.author | Susan Yun Fan Lin | en |
dc.contributor.author | 林芸帆 | zh_TW |
dc.date.accessioned | 2021-06-07T23:48:37Z | - |
dc.date.copyright | 2014-03-09 | |
dc.date.issued | 2014 | |
dc.date.submitted | 2014-02-17 | |
dc.identifier.citation | [1] Hanahan D, Weinberg RA. The hallmarks of cancer. Cell. 2000;100:57-70.
[2] Lazebnik Y. What are the hallmarks of cancer? Nature reviews Cancer. 2010;10:232-3. [3] Hutmacher DW LD, Rizzi S, Kaplan DL, Mooney DJ, Clements JA. Can tissue engineering concepts advance tumor biology research? Trends in Biotechnology. 2010;28:125-33. [4] Griffith LG, Swartz MA. Capturing complex 3D tissue physiology in vitro. Nature reviews Molecular cell biology. 2006;7:211-24. [5] Hutmacher DW. Biomaterials offer cancer research the third dimension. Nature materials. 2010;9:90-3. [6] Debnath J, Brugge JS. Modelling glandular epithelial cancers in three-dimensional cultures. Nature reviews Cancer. 2005;5:675-88. [7] Breslin S, O'Driscoll L. Three-dimensional cell culture: the missing link in drug discovery. Drug discovery today. 2013;18:240-9. [8] Gurski LA, Jha AK, Zhang C, Jia X, Farach-Carson MC. Hyaluronic acid-based hydrogels as 3D matrices for in vitro evaluation of chemotherapeutic drugs using poorly adherent prostate cancer cells. Biomaterials. 2009;30:6076-85. [9] Horning JL, Sahoo SK, Vijayaraghavalu S, Dimitrijevic S, Vasir JK, Jain TK, et al. 3-D tumor model for in vitro evaluation of anticancer drugs. Molecular pharmaceutics. 2008;5:849-62. [10] Hongisto V, Jernstrom S, Fey V, Mpindi JP, Kleivi Sahlberg K, Kallioniemi O, et al. High-throughput 3D screening reveals differences in drug sensitivities between culture models of JIMT1 breast cancer cells. PloS one. 2013;8:e77232. [11] Fischbach C, Chen R, Matsumoto T, Schmelzle T, Brugge JS, Polverini PJ, et al. Engineering tumors with 3D scaffolds. Nature methods. 2007;4:855-60. [12] Pampaloni F, Reynaud EG, Stelzer EH. The third dimension bridges the gap between cell culture and live tissue. Nature reviews Molecular cell biology. 2007;8:839-45. [13] Yamada KM, Cukierman E. Modeling tissue morphogenesis and cancer in 3D. Cell. 2007;130:601-10. [14] Fennema E, Rivron N, Rouwkema J, van Blitterswijk C, de Boer J. Spheroid culture as a tool for creating 3D complex tissues. Trends Biotechnol. 2013;31:108-15. [15] Kim JB. Three-dimensional tissue culture models in cancer biology. Seminars in cancer biology. 2005;15:365-77. [16] Su G, Zhao Y, Wei J, Han J, Chen L, Xiao Z, et al. The effect of forced growth of cells into 3D spheres using low attachment surfaces on the acquisition of stemness properties. Biomaterials. 2013;34:3215-22. [17] Chen SF, Chang YC, Nieh S, Liu CL, Yang CY, Lin YS. Nonadhesive culture system as a model of rapid sphere formation with cancer stem cell properties. PloS one. 2012;7:e31864. [18] Manuel Iglesias J, Beloqui I, Garcia-Garcia F, Leis O, Vazquez-Martin A, Eguiara A, et al. Mammosphere formation in breast carcinoma cell lines depends upon expression of E-cadherin. PloS one. 2013;8:e77281. [19] Ong SM, Zhao Z, Arooz T, Zhao D, Zhang S, Du T, et al. Engineering a scaffold-free 3D tumor model for in vitro drug penetration studies. Biomaterials. 2010;31:1180-90. [20] Fallica B, Makin G, Zaman MH. Bioengineering approaches to study multidrug resistance in tumor cells. Integrative biology : quantitative biosciences from nano to macro. 2011;3:529-39. [21] Godugu C, Patel AR, Desai U, Andey T, Sams A, Singh M. AlgiMatrix based 3D cell culture system as an in-vitro tumor model for anticancer studies. PloS one. 2013;8:e53708. [22] Dhiman HK, Ray AR, Panda AK. Three-dimensional chitosan scaffold-based MCF-7 cell culture for the determination of the cytotoxicity of tamoxifen. Biomaterials. 2005;26:979-86. [23] Florczyk SJ, Wang K, Jana S, Wood DL, Sytsma SK, Sham JG, et al. Porous chitosan-hyaluronic acid scaffolds as a mimic of glioblastoma microenvironment ECM. Biomaterials. 2013;34:10143-50. [24] Pedron S, Becka E, Harley BA. Regulation of glioma cell phenotype in 3D matrices by hyaluronic acid. Biomaterials. 2013;34:7408-17. [25] Xu X, Gurski LA, Zhang C, Harrington DA, Farach-Carson MC, Jia X. Recreating the tumor microenvironment in a bilayer, hyaluronic acid hydrogel construct for the growth of prostate cancer spheroids. Biomaterials. 2012;33:9049-60. [26] Ananthanarayanan B, Kim Y, Kumar S. Elucidating the mechanobiology of malignant brain tumors using a brain matrix-mimetic hyaluronic acid hydrogel platform. Biomaterials. 2011;32:7913-23. [27] Kang SW, Bae YH. Cryopreservable and tumorigenic three-dimensional tumor culture in porous poly(lactic-co-glycolic acid) microsphere. Biomaterials. 2009;30:4227-32. [28] Sahoo SK, Panda AK, Labhasetwar V. Characterization of porous PLGA/PLA microparticles as a scaffold for three dimensional growth of breast cancer cells. Biomacromolecules. 2005;6:1132-9. [29] Girard YK, Wang C, Ravi S, Howell MC, Mallela J, Alibrahim M, et al. A 3D fibrous scaffold inducing tumoroids: a platform for anticancer drug development. PloS one. 2013;8:e75345. [30] Chevallay B, Herbage D. Collagen-based biomaterials as 3D scaffold for cell cultures: applications for tissue engineering and gene therapy. Medical & biological engineering & computing. 2000;38:211-8. [31] Chen L, Xiao Z, Meng Y, Zhao Y, Han J, Su G, et al. The enhancement of cancer stem cell properties of MCF-7 cells in 3D collagen scaffolds for modeling of cancer and anti-cancer drugs. Biomaterials. 2012;33:1437-44. [32] Kirkland SC. Type I collagen inhibits differentiation and promotes a stem cell-like phenotype in human colorectal carcinoma cells. British journal of cancer. 2009;101:320-6. [33] Egeblad M, Rasch MG, Weaver VM. Dynamic interplay between the collagen scaffold and tumor evolution. Current opinion in cell biology. 2010;22:697-706. [34] Singh A, Settleman J. EMT, cancer stem cells and drug resistance: an emerging axis of evil in the war on cancer. Oncogene. 2010;29:4741-51. [35] Liang Y, Jeong J, DeVolder RJ, Cha C, Wang F, Tong YW, et al. A cell-instructive hydrogel to regulate malignancy of 3D tumor spheroids with matrix rigidity. Biomaterials. 2011;32:9308-15. [36] Yang YL, Motte S, Kaufman LJ. Pore size variable type I collagen gels and their interaction with glioma cells. Biomaterials. 2010;31:5678-88. [37] Harjanto D, Maffei JS, Zaman MH. Quantitative analysis of the effect of cancer invasiveness and collagen concentration on 3D matrix remodeling. PloS one. 2011;6:e24891. [38] Miron-Mendoza M, Seemann J, Grinnell F. The differential regulation of cell motile activity through matrix stiffness and porosity in three dimensional collagen matrices. Biomaterials. 2010;31:6425-35. [39] Wolf K, Alexander S, Schacht V, Coussens LM, von Andrian UH, van Rheenen J, et al. Collagen-based cell migration models in vitro and in vivo. Seminars in cell & developmental biology. 2009;20:931-41. [40] Gobeaux F, Mosser G, Anglo A, Panine P, Davidson P, Giraud-Guille MM, et al. Fibrillogenesis in dense collagen solutions: a physicochemical study. Journal of molecular biology. 2008;376:1509-22. [41] Bott K, Upton Z, Schrobback K, Ehrbar M, Hubbell JA, Lutolf MP, et al. The effect of matrix characteristics on fibroblast proliferation in 3D gels. Biomaterials. 2010;31:8454-64. [42] Serpooshan V, Julien M, Nguyen O, Wang H, Li A, Muja N, et al. Reduced hydraulic permeability of three-dimensional collagen scaffolds attenuates gel contraction and promotes the growth and differentiation of mesenchymal stem cells. Acta Biomater. 2010;6:3978-87. [43] Raub CB, Suresh V, Krasieva T, Lyubovitsky J, Mih JD, Putnam AJ, et al. Noninvasive assessment of collagen gel microstructure and mechanics using multiphoton microscopy. Biophysical journal. 2007;92:2212-22. [44] Szot CS, Buchanan CF, Freeman JW, Rylander MN. 3D in vitro bioengineered tumors based on collagen I hydrogels. Biomaterials. 2011;32:7905-12. [45] Nyga A, Loizidou M, Emberton M, Cheema U. A novel tissue engineered three-dimensional in vitro colorectal cancer model. Acta Biomater. 2013;9:7917-26. [46] Reya T, Morrison SJ, Clarke MF, Weissman IL. Stem cells, cancer, and cancer stem cells. Nature. 2001;414:105-11. [47] Elsaba TM, Martinez-Pomares L, Robins AR, Crook S, Seth R, Jackson D, et al. The stem cell marker CD133 associates with enhanced colony formation and cell motility in colorectal cancer. PloS one. 2010;5:e10714. [48] Todaro M, Alea MP, Di Stefano AB, Cammareri P, Vermeulen L, Iovino F, et al. Colon cancer stem cells dictate tumor growth and resist cell death by production of interleukin-4. Cell stem cell. 2007;1:389-402. [49] Chandrasekaran S, DeLouise LA. Enriching and characterizing cancer stem cell sub-populations in the WM115 melanoma cell line. Biomaterials. 2011;32:9316-27. [50] Gilbertson RJ, Graham TA. Cancer: Resolving the stem-cell debate. Nature. 2012;488:462-3. [51] Qiu X, Wang Z, Li Y, Miao Y, Ren Y, Luan Y. Characterization of sphere-forming cells with stem-like properties from the small cell lung cancer cell line H446. Cancer letters. 2012;323:161-70. [52] Liu WD, Zhang T, Wang CL, Meng HM, Song YW, Zhao Z, et al. Sphere-forming tumor cells possess stem-like properties in human fibrosarcoma primary tumors and cell lines. Oncology letters. 2012;4:1315-20. [53] Han XY, Wei B, Fang JF, Zhang S, Zhang FC, Zhang HB, et al. Epithelial-mesenchymal transition associates with maintenance of stemness in spheroid-derived stem-like colon cancer cells. PloS one. 2013;8:e73341. [54] Mani SA, Guo W, Liao MJ, Eaton EN, Ayyanan A, Zhou AY, et al. The epithelial-mesenchymal transition generates cells with properties of stem cells. Cell. 2008;133:704-15. [55] Polyak K, Weinberg RA. Transitions between epithelial and mesenchymal states: acquisition of malignant and stem cell traits. Nature Reviews Cancer. 2009;9:265-73. [56] Christiansen JJ, Rajasekaran AK. Reassessing epithelial to mesenchymal transition as a prerequisite for carcinoma invasion and metastasis. Cancer research. 2006;66:8319-26. [57] Zeisberg M, Neilson EG. Biomarkers for epithelial-mesenchymal transitions. The Journal of clinical investigation. 2009;119:1429-37. [58] Chandrasekaran S, Geng Y, DeLouise LA, King MR. Effect of homotypic and heterotypic interaction in 3D on the E-selectin mediated adhesive properties of breast cancer cell lines. Biomaterials. 2012;33:9037-48. [59] Koenig A, Mueller C, Hasel C, Adler G, Menke A. Collagen type I induces disruption of E-cadherin-mediated cell-cell contacts and promotes proliferation of pancreatic carcinoma cells. Cancer research. 2006;66:4662-71. [60] Cheng JC, Leung PC. Type I collagen down-regulates E-cadherin expression by increasing PI3KCA in cancer cells. Cancer letters. 2011;304:107-16. [61] Imamichi Y, Konig A, Gress T, Menke A. Collagen type I-induced Smad-interacting protein 1 expression downregulates E-cadherin in pancreatic cancer. Oncogene. 2007;26:2381-5. [62] Carmeliet P, Jain RK. Angiogenesis in cancer and other diseases. Nature. 2000;407:249-57. [63] Liotta LA, Steeg PS, Stetler-Stevenson WG. Cancer metastasis and angiogenesis: an imbalance of positive and negative regulation. Cell. 1991;64:327-36. [64] Fidler IJ, Ellis LM. The implications of angiogenesis for the biology and therapy of cancer metastasis. Cell. 1994;79:185-8. [65] Carmeliet P. VEGF as a key mediator of angiogenesis in cancer. Oncology. 2005;69 Suppl 3:4-10. [66] Martiny-Baron G, Marme D. VEGF-mediated tumour angiogenesis: a new target for cancer therapy. Current opinion in biotechnology. 1995;6:675-80. [67] Zhou J, Schmid T, Schnitzer S, Brune B. Tumor hypoxia and cancer progression. Cancer letters. 2006;237:10-21. [68] Drug resistance in cancer chemotherapy. Nature. 1971;233:162. [69] Minchinton AI, Tannock IF. Drug penetration in solid tumours. Nature reviews Cancer. 2006;6:583-92. [70] A. A, MB. S. The tumour microenvironment as a target for chemoprevention. Nature reviews Cancer. 2007;7:139-47. [71] Tannock IF, Lee CM, Tunggal JK, Cowan DS, Egorin MJ. Limited penetration of anticancer drugs through tumor tissue: a potential cause of resistance of solid tumors to chemotherapy. Clinical cancer research : an official journal of the American Association for Cancer Research. 2002;8:878-84. [72] Seo BR, Delnero P, Fischbach C. In vitro models of tumor vessels and matrix: Engineering approaches to investigate transport limitations and drug delivery in cancer. Advanced drug delivery reviews. 2013. [73] Tredan O, Galmarini CM, Patel K, Tannock IF. Drug resistance and the solid tumor microenvironment. Journal of the National Cancer Institute. 2007;99:1441-54. [74] Loessner D, Stok KS, Lutolf MP, Hutmacher DW, Clements JA, Rizzi SC. Bioengineered 3D platform to explore cell-ECM interactions and drug resistance of epithelial ovarian cancer cells. Biomaterials. 2010;31:8494-506. [75] Leeper AD, Farrell J, Williams LJ, Thomas JS, Dixon JM, Wedden SE, et al. Determining tamoxifen sensitivity using primary breast cancer tissue in collagen-based three-dimensional culture. Biomaterials. 2012;33:907-15. [76] Moro L, Arbini AA, Marra E, Greco M. Down-regulation of BRCA2 expression by collagen type I promotes prostate cancer cell proliferation. The Journal of biological chemistry. 2005;280:22482-91. [77] Shintani Y, Maeda M, Chaika N, Johnson KR, Wheelock MJ. Collagen I promotes epithelial-to-mesenchymal transition in lung cancer cells via transforming growth factor-beta signaling. American journal of respiratory cell and molecular biology. 2008;38:95-104. [78] Cross VL, Zheng Y, Won Choi N, Verbridge SS, Sutermaster BA, Bonassar LJ, et al. Dense type I collagen matrices that support cellular remodeling and microfabrication for studies of tumor angiogenesis and vasculogenesis in vitro. Biomaterials. 2010;31:8596-607. [79] Furukawa T, Kubota T, Watanabe M, Kase S, Saikawa Y, Nishibori H, et al. Increased drug resistance of cultured human cancer cell lines in three-dimensional cellular growth assay using collagen gel matrix. Journal of surgical oncology. 1992;49:86-92. [80] Paszek MJ, Zahir N, Johnson KR, Lakins JN, Rozenberg GI, Gefen A, et al. Tensional homeostasis and the malignant phenotype. Cancer cell. 2005;8:241-54. [81] Levental KR, Yu H, Kass L, Lakins JN, Egeblad M, Erler JT, et al. Matrix crosslinking forces tumor progression by enhancing integrin signaling. Cell. 2009;139:891-906. [82] Pozzi A, Wary KK, Giancotti FG, Gardner HA. Integrin alpha1beta1 mediates a unique collagen-dependent proliferation pathway in vivo. The Journal of cell biology. 1998;142:587-94. [83] Fischbach C, Kong HJ, Hsiong SX, Evangelista MB, Yuen W, Mooney DJ. Cancer cell angiogenic capability is regulated by 3D culture and integrin engagement. Proc Natl Acad Sci U S A. 2009;106:399-404. [84] Hall CL, Dai J, van Golen KL, Keller ET, Long MW. Type I collagen receptor (alpha 2 beta 1) signaling promotes the growth of human prostate cancer cells within the bone. Cancer research. 2006;66:8648-54. [85] Hall CL, Dubyk CW, Riesenberger TA, Shein D, Keller ET, van Golen KL. Type I collagen receptor (alpha2beta1) signaling promotes prostate cancer invasion through RhoC GTPase. Neoplasia. 2008;10:797-803. [86] Dolznig H, Rupp C, Puri C, Haslinger C, Schweifer N, Wieser E, et al. Modeling colon adenocarcinomas in vitro a 3D co-culture system induces cancer-relevant pathways upon tumor cell and stromal fibroblast interaction. The American journal of pathology. 2011;179:487-501. [87] Windus LC, Glover TT, Avery VM. Bone-stromal cells up-regulate tumourigenic markers in a tumour-stromal 3D model of prostate cancer. Molecular cancer. 2013;12:112. [88] Fang X, Sittadjody S, Gyabaah K, Opara EC, Balaji KC. Novel 3D co-culture model for epithelial-stromal cells interaction in prostate cancer. PloS one. 2013;8:e75187. [89] Sung KE, Su X, Berthier E, Pehlke C, Friedl A, Beebe DJ. Understanding the impact of 2D and 3D fibroblast cultures on in vitro breast cancer models. PloS one. 2013;8:e76373. [90] Correa de Sampaio P, Auslaender D, Krubasik D, Failla AV, Skepper JN, Murphy G, et al. A heterogeneous in vitro three dimensional model of tumour-stroma interactions regulating sprouting angiogenesis. PloS one. 2012;7:e30753. [91] Yang YL, Sun C, Wilhelm ME, Fox LJ, Zhu J, Kaufman LJ. Influence of chondroitin sulfate and hyaluronic acid on structure, mechanical properties, and glioma invasion of collagen I gels. Biomaterials. 2011;32:7932-40. [92] Wang L, Murthy SK, Barabino GA, Carrier RL. Synergic effects of crypt-like topography and ECM proteins on intestinal cell behavior in collagen based membranes. Biomaterials. 2010;31:7586-98. [93] Banerjee SS, Paul D, Bhansali SG, Aher ND, Jalota-Badhwar A, Khandare J. Enhancing surface interactions with colon cancer cells on a transferrin-conjugated 3D nanostructured substrate. Small. 2012;8:1657-63. [94] Butcher DT, Alliston T, Weaver VM. A tense situation: forcing tumour progression. Nature reviews Cancer. 2009;9:108-22. [95] Cukierman E, Bassi DE. Physico-mechanical aspects of extracellular matrix influences on tumorigenic behaviors. Seminars in cancer biology. 2010;20:139-45. [96] Beck JN, Singh A, Rothenberg AR, Elisseeff JH, Ewald AJ. The independent roles of mechanical, structural and adhesion characteristics of 3D hydrogels on the regulation of cancer invasion and dissemination. Biomaterials. 2013;34:9486-95. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/16876 | - |
dc.description.abstract | 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。 | zh_TW |
dc.description.abstract | 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. | en |
dc.description.provenance | Made available in DSpace on 2021-06-07T23:48:37Z (GMT). No. of bitstreams: 1 ntu-103-R00548063-1.pdf: 3348143 bytes, checksum: 06d5fc8b686dd8558195a98ca4022219 (MD5) Previous issue date: 2014 | en |
dc.description.tableofcontents | CONTENTS
摘要 i ABSTRACT ii CONTENTS iv LISTS OF SCHEMES viii LIST OF TABLES ix LIST OF FIGURES x Chapter 1 Introduction 1 Chapter 2 Materials and Methods 4 2.1 Materials 4 2.2 Cell culture 4 2.3 2D monolayer model development 4 2.4 3D spheroid model development 5 2.5 3D gel/scaffold-embedding model development 5 2.6 Optimization of type I collagen gel formulation for 3D in vitro gel/scaffold-embedding model development of human colorectal cancer cells 6 2.7 Rheometry 6 2.8 Inverted light microscopy 7 2.9 Cell harvesting 7 2.10 Trypan blue exclusion assay and cell counting 8 2.11 Reverse transcription – real-time polymerase chain reaction (RT-qPCR) 8 2.12 Western blot 8 2.13 Flow cytometry 9 2.14 Enzyme-linked immunosorbent assay (ELISA) 9 2.15 Cytotoxicity assay 10 2.16 In vivo tumorigenicity assay 10 2.17 Statistical analysis 10 Chapter 3 Results 12 3.1 Search for the optimal type I collagen gel formulation for 3D in vitro gel/scaffold- embedding model development of human colorectal cancer cells. 12 3.2 Cell morphology comparison among the three in vitro colorectal cancer cell models. 13 3.3 Cell viability comparison among the three in vitro colorectal cancer cell models. 13 3.4 Cancer stem cell marker expression comparison among the three in vitro colorectal cancer cell models. 14 3.5 Stemness markers expression comparison among the three in vitro colorectal cancer cell models. 14 3.6 Epithelial-mesenchymal transition markers expression comparison among the three in vitro colorectal cancer cell models. 15 3.7 Angiogenesis marker expression comparison among the three in vitro colorectal cancer cell models. 16 3.8 Anti-cancer drug resistance comparison among the three in vitro colorectal cancer cell models. 17 3.9 In vivo tumorigenicity comparison among the three in vitro colorectal cancer cell models. 17 Chapter 4 Discussion 19 4.1 The optimal type I collagen gel formulation for 3D in vitro gel/scaffold-embedding model development of human colorectal cancer cells. 19 4.2 Reproducibility of in vivo cancer cell morphology is the highest in 3D cancer cell spheres cultured within type I collagen gel. 21 4.3 Cell viability is the highest in 3D cancer cell spheres cultured within type I collagen gel. 22 4.4 Colorectal cancer stem cell population is the highest in 3D cancer cell spheres cultured within type I collagen gel. 23 4.5 Stemness markers expression is the highest in 3D cancer cell spheres cultured within type I collagen gel. 24 4.6 Epithelial-mesenchymal transition markers expression is the highest in 3D cancer cell spheres cultured within type I collagen gel. 25 4.7 Angiogenesis marker expression is the highest in 3D cancer cell spheres cultured within type I collagen gel. 26 4.8 Anti-cancer drug resistance is the highest in 3D cancer cell spheres cultured within type I collagen gel. 27 4.9 In vivo tumorigenicity is the highest in 3D cancer cell spheres cultured within type I collagen gel. 28 4.10 The bias of result by type I collagen gel. 29 4.11 3D muticellularity and microenvironment are the basic constituents toward the development of a biomimetic 3D in vitro cancer cell model. 29 Chapter 5 Conclusion 31 REFERENCE 32 SCHEME 43 TABLE 45 FIGURE 46 | |
dc.language.iso | zh-TW | |
dc.title | 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 | zh_TW |
dc.title | The Development of a Biomimetic In Vitro Model with 3D Spheres of Human Colorectal Cancer Cells embedded within Type I Collagen Gel | en |
dc.type | Thesis | |
dc.date.schoolyear | 102-1 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 楊台鴻,婁培人 | |
dc.subject.keyword | 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, | zh_TW |
dc.subject.keyword | 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, | en |
dc.relation.page | 77 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2014-02-17 | |
dc.contributor.author-college | 工學院 | zh_TW |
dc.contributor.author-dept | 醫學工程學研究所 | zh_TW |
顯示於系所單位: | 醫學工程學研究所 |
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