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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 陳羿貞 | zh_TW |
dc.contributor.advisor | Yi-Jane Chan | en |
dc.contributor.author | 劉上維 | zh_TW |
dc.contributor.author | Shang-Wei Liu | en |
dc.date.accessioned | 2023-09-26T16:15:29Z | - |
dc.date.available | 2023-11-10 | - |
dc.date.copyright | 2023-09-26 | - |
dc.date.issued | 2023 | - |
dc.date.submitted | 2023-07-17 | - |
dc.identifier.citation | 1. Till, J. E. and Culloch Ea Mc (1961). "A direct measurement of the radiation sensitivity of normal mouse bone marrow cells." Radiation Research 14: 213-222.
2. Gronthos, S., M. Mankani, J. Brahim, P. G. Robey and S. Shi (2000). "Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo." Proceedings of the National Academy of Sciences of the United States of America 97(25): 13625-13630. 3. Lan, X. Y., Z. W. Sun, C. Y. Chu, J. Boltze and S. Li (2019). "Dental Pulp Stem Cells: An Attractive Alternative for Cell Therapy in Ischemic Stroke." Frontiers in Neurology 10. 4. Pittenger, M. F., D. E. Discher, B. M. Péault, D. G. Phinney, J. M. Hare and A. I. Caplan (2019). "Mesenchymal stem cell perspective: cell biology to clinical progress." NPJ Regenerative Medicine 4: 22. 5. Bronckaers, A., P. Hilkens, Y. Fanton, T. Struys, P. Gervois, C. Politis, W. Martens and I. Lambrichts (2013). "Angiogenic properties of human dental pulp stem cells." PLoS One 8(8): e71104. 6. Knight, E. and S. Przyborski (2015). "Advances in 3D cell culture technologies enabling tissue-like structures to be created in vitro." Journal of Anatomy 227(6): 746-756. 7. Vinci, M., S. Gowan, F. Boxall, L. Patterson, M. Zimmermann, W. Court, C. Lomas, M. Mendiola, D. Hardisson and S. A. Eccles (2012). "Advances in establishment and analysis of three-dimensional tumor spheroid-based functional assays for target validation and drug evaluation." BMC Biology 10: 29. 8. Lv, D., Z. Hu, L. Lu, H. Lu and X. Xu (2017). "Three-dimensional cell culture: A powerful tool in tumor research and drug discovery." Oncology Letters 14(6): 6999-7010. 9. Cesarz, Z. and K. Tamama (2016). "Spheroid Culture of Mesenchymal Stem Cells." Stem Cells International 2016: 9176357. 10. Fang, Y. and R. M. Eglen (2017). "Three-Dimensional Cell Cultures in Drug Discovery and Development." SLAS Discovery 22(5): 456-472. 11. Ryu, N. E., S. H. Lee and H. Park (2019). "Spheroid Culture System Methods and Applications for Mesenchymal Stem Cells." Cells 8(12). 12. Petrenko, Y., E. Sykova and S. Kubinova (2017). "The therapeutic potential of three-dimensional multipotent mesenchymal stromal cell spheroids." Stem Cell Research & Therapy 8. 13. Chen, C. S., M. Mrksich, S. Huang, G. M. Whitesides and D. E. Ingber (1997). "Geometric control of cell life and death." Science 276(5317): 1425-1428. 14. Edmondson, R., J. J. Broglie, A. F. Adcock and L. Yang (2014). "Three-dimensional cell culture systems and their applications in drug discovery and cell-based biosensors." ASSAY and Drug Development Technologies 12(4): 207-218. 15. Singhvi, R., A. Kumar, G. P. Lopez, G. N. Stephanopoulos, D. I. Wang, G. M. Whitesides and D. E. Ingber (1994). "Engineering cell shape and function." Science 264(5159): 696-698. 16. Khaitan, D., S. Chandna, M. B. Arya and B. S. Dwarakanath (2006). "Establishment and characterization of multicellular spheroids from a human glioma cell line; Implications for tumor therapy." Journal of Translational Medicine 4: 12. 17. Kim, J. B. (2005). "Three-dimensional tissue culture models in cancer biology." Seminars in Cancer Biology 15(5): 365-377. 18. Kenny, P. A., G. Y. Lee, C. A. Myers, R. M. Neve, J. R. Semeiks, P. T. Spellman, K. Lorenz, E. H. Lee, M. H. Barcellos-Hoff, O. W. Petersen, J. W. Gray and M. J. Bissell (2007). "The morphologies of breast cancer cell lines in three-dimensional assays correlate with their profiles of gene expression." Molecular Oncology 1(1): 84-96. 19. Lin, R. Z. and H. Y. Chang (2008). "Recent advances in three-dimensional multicellular spheroid culture for biomedical research." Biotechnology Journal 3(9-10): 1172-1184. 20. Schmitz, C., E. Potekhina, V. V. Belousov and A. Lavrentieva (2021). "Hypoxia Onset in Mesenchymal Stem Cell Spheroids: Monitoring With Hypoxia Reporter Cells." Frontiers in Bioengineering and Biotechnology 9. 21. Hsieh, H. Y., T. H. Young, C. C. Yao and Y. J. Chen (2019). "Aggregation of human dental pulp cells into 3D spheroids enhances their migration ability after reseeding." Journal of Cellular Physiology 234(1): 976-986. 22. Tamama, K., H. Kawasaki, S. S. Kerpedjieva, J. Guan, R. K. Ganju and C. K. Sen (2011). "Differential roles of hypoxia inducible factor subunits in multipotential stromal cells under hypoxic condition." Journal of Cellular Biochemistry 112(3): 804-817. 23. Wan, C., J. Shao, S. R. Gilbert, R. C. Riddle, F. Long, R. S. Johnson, E. Schipani and T. L. Clemens (2010). "Role of HIF-1alpha in skeletal development." Annals of the New York Academy of Sciences journal 1192: 322-326. 24. Morimoto, M., H. Saimoto, H. Usui, Y. Okamoto, S. Minami and Y. Shigemasa (2001). "Biological activities of carbohydrate-branched chitosan derivatives." Biomacromolecules 2(4): 1133-1136. 25. Shi, C., Y. Zhu, X. Ran, M. Wang, Y. Su and T. Cheng (2006). "Therapeutic potential of chitosan and its derivatives in regenerative medicine." Journal of Surgical Research 133(2): 185-192. 26. Shigemasa, Y. and S. Minami (1996). "Applications of chitin and chitosan for biomaterials." Biotechnology and Genetic Engineering Reviews 13: 383-420. 27. Hsu, S. H., Lin, T. L., Chiang, L. T., Tseng, T. C. Lee, H. T.,Liao, C. Y., and Chiu, I. M. (2012). "Spheroid Formation from Neural Stem Cells on Chitosan Membranes." Journal of Medical and Biological Engineering 32. 28. Hsu, S. H., Huang, G. S. and Feng, F. (2012). "Isolation of the multipotent MSC subpopulation from human gingival fibroblasts by culturing on chitosan membranes." Biomaterials 33(9): 2642-2655. 29. Yeh, H. Y., B. H. Liu and S. H. Hsu (2012). "The calcium-dependent regulation of spheroid formation and cardiomyogenic differentiation for MSCs on chitosan membranes." Biomaterials 33(35): 8943-8954. 30. Huang, G. S., L. G. Dai, B. L. Yen and S. H. Hsu (2011). "Spheroid formation of mesenchymal stem cells on chitosan and chitosan-hyaluronan membranes." Biomaterials 32(29): 6929-6945. 31. Yeh, H. Y., B. H. Liu, M. Sieber and S. H. Hsu (2014). "Substrate-dependent gene regulation of self-assembled human MSC spheroids on chitosan membranes." BMC Genomics 15(1): 10. 32. Cheng, N. C., S. Wang and T. H. Young (2012). "The influence of spheroid formation of human adipose-derived stem cells on chitosan films on stemness and differentiation capabilities." Biomaterials 33(6): 1748-1758. 33. Chen, Y. H., I. J. Wang and T. H. Young (2009). "Formation of keratocyte spheroids on chitosan-coated surface can maintain keratocyte phenotypes." Tissue Engineering Part A 15(8): 2001-2013. 34. Lin, S. J., S. H. Jee, W. C. Hsaio, S. J. Lee and T. H. Young (2005). "Formation of melanocyte spheroids on the chitosan-coated surface." Biomaterials 26(12): 1413-1422. 35. Murphy, K. C., J. Whitehead, P. C. Falahee, D. Zhou, S. I. Simon and J. K. Leach (2017). "Multifactorial Experimental Design to Optimize the Anti-Inflammatory and Proangiogenic Potential of Mesenchymal Stem Cell Spheroids." Stem Cells 35(6): 1493-1504. 36. Yan, X-Z, JJJP van den Beucken, C Yuan, JA Jansen and F Yang (2018). "Spheroid formation and stemness preservation of human periodontal ligament cells on chitosan films." Oral Diseases 24(6): 1083-1092. 37. Carmeliet, P. (2005). "Angiogenesis in life, disease and medicine." Nature 438(7070): 932-936. 38. Adair, T. H. and J. P. Montani (2010). Integrated Systems Physiology: from Molecule to Function to Disease. Angiogenesis. San Rafael (CA), Morgan & Claypool Life Sciences Copyright © 2010 by Morgan & Claypool Life Sciences. 39. Folkman, Judah (2008). History of Angiogenesis. Angiogenesis: An Integrative Approach From Science to Medicine. William D. Figg and Judah Folkman. Boston, MA, Springer US: 1-14. 40. Greene, A. K., S. Wiener, M. Puder, A. Yoshida, B. Shi, A. R. Perez-Atayde, J. A. Efstathiou, L. Holmgren, A. P. Adamis, M. Rupnick, J. Folkman and M. S. O'Reilly (2003). "Endothelial-directed hepatic regeneration after partial hepatectomy." Annals of Surgery 237(4): 530-535. 41. Farooq, Mariya, Abdul Waheed Khan, Moon Suk Kim and Sangdun Choi (2021). "The Role of Fibroblast Growth Factor (FGF) Signaling in Tissue Repair and Regeneration." Cells 10(11): 3242. 42. Shibuya, Masabumi (2008). Vascular Permeability/Vascular Endothelial Growth Factor. Angiogenesis: An Integrative Approach From Science to Medicine. William D. Figg and Judah Folkman. Boston, MA, Springer US: 89-98. 43. Nakamichi, M., Y. Akishima-Fukasawa, C. Fujisawa, T. Mikami, K. Onishi and Y. Akasaka (2016). "Basic Fibroblast Growth Factor Induces Angiogenic Properties of Fibrocytes to Stimulate Vascular Formation during Wound Healing." The American Journal of Pathology 186(12): 3203-3216. 44. Gao, X. and Z. Xu (2008). "Mechanisms of action of angiogenin." Acta Biochimica et Biophysica Sinica 40(7): 619-624. 45. Hoang, T. T. and R. T. Raines (2016). "Molecular basis for the autonomous promotion of cell proliferation by angiogenin." Nucleic Acids Research 45(2): 818-831. 46. Brindle, N. P., P. Saharinen and K. Alitalo (2006). "Signaling and functions of angiopoietin-1 in vascular protection." Circulation Research 98(8): 1014-1023. 47. Lin, T. N., C. K. Wang, W. M. Cheung and C. Y. Hsu (2000). "Induction of Angiopoietin and Tie Receptor mRNA Expression after Cerebral Ischemia–Reperfusion." Journal of Cerebral Blood Flow & Metabolism 20(2): 387-395. 48. Akwii, Racheal G., Md S. Sajib, Fatema T. Zahra and Constantinos M. Mikelis (2019). "Role of Angiopoietin-2 in Vascular Physiology and Pathophysiology." Cells 8(5): 471. 49. Krock, B. L., N. Skuli and M. C. Simon (2011). "Hypoxia-induced angiogenesis: good and evil." Genes Cancer 2(12): 1117-1133. 50. Pugh, C. W. and P. J. Ratcliffe (2003). "Regulation of angiogenesis by hypoxia: role of the HIF system." Nature Medicine 9(6): 677-684. 51. Ge, L., C. F. Xun, W. S. Li, S. Y. Jin, Z. Liu, Y. Zhuo, D. Duan, Z. P. Hu, P. Chen and M. Lu (2021). "Extracellular vesicles derived from hypoxia-preconditioned olfactory mucosa mesenchymal stem cells enhance angiogenesis via miR-612." Journal of Nanobiotechnology 19(1): 380. 52. Han, Y. D., J. Ren, Y. Bai, X. T. Pei and Y. Han (2019). "Exosomes from hypoxia-treated human adipose-derived mesenchymal stem cells enhance angiogenesis through VEGF/VEGF-R." The International Journal of Biochemistry & Cell Biology 109: 59-68. 53. Gregorius, J., C. Wang, O. Stambouli, T. Hussner, Y. Qi, T. Tertel, V. Börger, A. M. Yusuf, N. Hagemann, D. P. Yin, R. Dittrich, Y. Mouloud, F. D. Mairinger, F. El Magraoui, A. Popa-Wagner, C. Kleinschnitz, T. R. Doeppner, M. Gunzer, H. E. Meyer, B. Giebel and D. M. Hermann (2021). "Small extracellular vesicles obtained from hypoxic mesenchymal stromal cells have unique characteristics that promote cerebral angiogenesis, brain remodeling and neurological recovery after focal cerebral ischemia in mice." Basic Research in Cardiology 116(1): 40. 54. Hu, N., Z. W. Cai, X. D. Jiang, C. Wang, T. Tang, T. Xu, H. Chen, X. Q. Li, X.L. Du and W. G., Cui (2023). "Hypoxia-pretreated ADSC-derived exosome-embedded hydrogels promote angiogenesis and accelerate diabetic wound healing." Acta Biomaterialia 157: 175-186. 55. Antonio Nanci. (2013) Chapter 8 - Dentin-Pulp Complex. Ten Cate's Oral Histology (Eighth Edition.). St. Louis (MO), Mosby: 165-204. 56. Nuti, N., C. Corallo, B. M. F. Chan, M. Ferrari and B. Gerami-Naini (2016). "Multipotent Differentiation of Human Dental Pulp Stem Cells: a Literature Review." Stem Cell Reviews and Reports 12(5): 511-523. 57. Nakashima, M., K. Iohara and M. Sugiyama (2009). "Human dental pulp stem cells with highly angiogenic and neurogenic potential for possible use in pulp regeneration." Cytokine Growth Factor Reviews 20(5-6): 435-440. 58. Gharaei, M. A., Y. Xue, K. Mustafa, S. A. Lie and I. Fristad (2018). "Human dental pulp stromal cell conditioned medium alters endothelial cell behavior." Stem Cell Research & Therapy 9. 59. Sieveking, D. P. and M. K. Ng (2009). "Cell therapies for therapeutic angiogenesis: back to the bench." Vascular Medicine 14(2): 153-166. 60. Aranha, A. M. F., Z. C. Zhang, K. G. Neiva, C. A. S. Costa, J. Hebling and J. E. Nör (2010). "Hypoxia Enhances the Angiogenic Potential of Human Dental Pulp Cells." Journal of Endodontics 36(10): 1633-1637. 61. Ishizaka, R., Y. Hayashi, K. Iohara, M. Sugiyama, M. Murakami, T. Yamamoto, O. Fukuta and M. Nakashima (2013). "Stimulation of angiogenesis, neurogenesis and regeneration by side population cells from dental pulp." Biomaterials 34(8): 1888-1897. 62. Gandia, C., A. Armiñan, J. M. García-Verdugo, E. Lledó, A. Ruiz, M. D. Miñana, J. Sanchez-Torrijos, R. Payá, V. Mirabet, F. Carbonell-Uberos, M. Llop, J. A. Montero and P. Sepúlveda (2008). "Human dental pulp stem cells improve left ventricular function, induce angiogenesis, and reduce infarct size in rats with acute myocardial infarction." Stem Cells 26(3): 638-645. 63. Dissanayaka, W. L., X. Zhan, C. F. Zhang, K. M. Hargreaves, L. J. Jin and E. H. Y. Tong (2012). "Coculture of Dental Pulp Stem Cells with Endothelial Cells Enhances Osteo-/Odontogenic and Angiogenic Potential In Vitro." Journal of Endodontics 38(4): 454-463. 64. Fujio, M., Z. Xing, N. Sharabi, Y. Xue, A. Yamamoto, H. Hibi, M. Ueda, I. Fristad and K. Mustafa (2017). "Conditioned media from hypoxic-cultured human dental pulp cells promotes bone healing during distraction osteogenesis." Journal of Tissue Engineering and Regenerative Medicine 11(7): 2116-2126. 65. Hirschhaeuser, F., H. Menne, C. Dittfeld, J. West, W. Mueller-Klieser and L. A. Kunz-Schughart (2010). "Multicellular tumor spheroids: an underestimated tool is catching up again." Journal of Biotechnology 148(1): 3-15. 66. Tietze, S. , M. Kräter, A. Jacobi, A. Taubenberger, M. Herbig, R. Wehner, M. Schmitz, O. Otto, C. List, B. Kaya, M. Wobus, M. Bornhäuser and J. Guck (2019). "Spheroid Culture of Mesenchymal Stromal Cells Results in Morphorheological Properties Appropriate for Improved Microcirculation." Advanced Science 6(8): 1802104. 67. Lautenschläger, F., S. Paschke, S. Schinkinger, A. Bruel, M. Beil and J. Guck (2009). "The regulatory role of cell mechanics for migration of differentiating myeloid cells." Proceedings of the National Academy of Sciences of the United States of America 106(37): 15696-15701. 68. Mo, M., Y. Zhou, S. Li and Y. Wu (2018). "Three-Dimensional Culture Reduces Cell Size By Increasing Vesicle Excretion." Stem Cells 36(2): 286-292. 69. Ge, J., L. Guo, S. Wang, Y. Zhang, T. Cai, R. C. Zhao and Y. Wu (2014). "The size of mesenchymal stem cells is a significant cause of vascular obstructions and stroke." Stem Cell Reviews and Reports 10(2): 295-303. 70. Bhang, S. H., S. Lee, J. Y. Shin, T. J. Lee, H. K. Jang and B. S. Kim (2014). "Efficacious and clinically relevant conditioned medium of human adipose-derived stem cells for therapeutic angiogenesis." Molecular Therapy 22(4): 862-872. 71. Tran-Hung, L., P. Laurent, J. Camps and I. About (2008). "Quantification of angiogenic growth factors released by human dental cells after injury." Archives of Oral Biology 53(1): 9-13. 72. Kifune, T., H. Ito, M. Ishiyama, S. Iwasa, H. Takei, T. Hasegawa, M. Asano and T. Shirakawa (2018). "Hypoxia-induced upregulation of angiogenic factors in immortalized human periodontal ligament fibroblasts." Journal of Oral Science 60(4): 519-525. 73. Xiong, X., Y. Sun and X. C. Wang (2020). "HIF1A/miR-20a-5p/TGFβ1 axis modulates adipose-derived stem cells in a paracrine manner to affect the angiogenesis of human dermal microvascular endothelial cells." Journal of Cellular Physiology 235(3): 2091-2101. 74. Paola, A. Guerrero and H. McCarty Joseph (2017). TGF-β Activation and Signaling in Angiogenesis. Physiologic and Pathologic Angiogenesis. Simionescu Dan and Simionescu Agneta. Rijeka, IntechOpen: Ch. 1. 75. Fang, L. L., Y. R. Li, S. J. Wang, Y. X. Li, H. M. Chang, Y. Y. Yi, Y. Yan, A. Thakur, P. C. K. Leung, J. C. Cheng and Y. P. Sun (2020). "TGF-β1 induces VEGF expression in human granulosa-lutein cells: a potential mechanism for the pathogenesis of ovarian hyperstimulation syndrome." Experimental & Molecular Medicine 52(3): 450-460. 76. Zheng, W., E. A. Seftor, C. J. Meininger, M. J. Hendrix and R. J. Tomanek (2001). "Mechanisms of coronary angiogenesis in response to stretch: role of VEGF and TGF-beta." American Journal of Physiology-Heart and Circulatory Physiology 280(2): H909-917. 77. Hayes, A. J., W. Q.Huang, J. Mallah, D. J. Yang, M. E. Lippman and L. Y. Li (1999). "Angiopoietin-1 and Its Receptor Tie-2 Participate in the Regulation of Capillary-like Tubule Formation and Survival of Endothelial Cells." Microvascular Research 58(3): 224-237. 78. Koh, G. Y. (2013). "Orchestral actions of angiopoietin-1 in vascular regeneration." Trends in Molecular Medicine 19(1): 31-39. 79. Holash, J., S. J. Wiegand and G. D. Yancopoulos (1999). "New model of tumor angiogenesis: dynamic balance between vessel regression and growth mediated by angiopoietins and VEGF." Oncogene 18(38): 5356-5362. 80. Rey, S. and G. L. Semenza (2010). "Hypoxia-inducible factor-1-dependent mechanisms of vascularization and vascular remodelling." Cardiovascular Research 86(2): 236-242. 81. Park, Y. S., N. H. Kim and I. Jo (2003). "Hypoxia and vascular endothelial growth factor acutely up-regulate angiopoietin-1 and Tie2 mRNA in bovine retinal pericytes." Microvascular Research 65(2): 125-131. 82. Lee, E. Y., Y. Xia, W. S. Kim, M. H. Kim, T. H. Kim, K. J. Kim, B. S. Park and J. H. Sung (2009). "Hypoxia-enhanced wound-healing function of adipose-derived stem cells: Increase in stem cell proliferation and up-regulation of VEGF and bFGF." Wound Repair and Regeneration 17(4): 540-547. 83. Calvani, M., A. Rapisarda, B. Uranchimeg, R. H. Shoemaker and G. Melillo (2006). "Hypoxic induction of an HIF-1alpha-dependent bFGF autocrine loop drives angiogenesis in human endothelial cells." Blood 107(7): 2705-2712. 84. Rao, Z. H., D. P. Shen, J. H. Chen, L. S. Jin, X. P. Wu, M. Chen, L. Li, M. P. Chu and J. F. Lin (2020). "Basic Fibroblast Growth Factor Attenuates Injury in Myocardial Infarction by Enhancing Hypoxia-Inducible Factor-1 Alpha Accumulation." Frontiers in Pharmacology 11. 85. Kishimoto, K., S. Yoshida, S. Ibaragi, N. Yoshioka, T. Okui, G. F. Hu and A. Sasaki (2012). "Hypoxia-induced up-regulation of angiogenin, besides VEGF, is related to progression of oral cancer." Oral Oncology 48(11): 1120-1127. 86. Nakamura, M., H. Yamabe, H. Osawa, N. Nakamura, M. Shimada, R. Kumasaka, R. Murakami, T. Fujita, T. Osanai and K. Okumura (2006). "Hypoxic conditions stimulate the production of angiogenin and vascular endothelial growth factor by human renal proximal tubular epithelial cells in culture." Nephrology Dialysis Transplantation 21(6): 1489-1495. 87. Janjić, K., M. Edelmayer, A. Moritz and H. Agis (2017). "L-mimosine and hypoxia can increase angiogenin production in dental pulp-derived cells." BMC Oral Health 17(1): 87. 88. Valadi, H., K. Ekström, A. Bossios, M. Sjöstrand, J. J. Lee and J. O. Lötvall (2007). "Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells." Nature Cell Biology 9(6): 654-659. 89. Nakamura, Y., S. Miyaki, H. Ishitobi, S. Matsuyama, T. Nakasa, N. Kamei, T. Akimoto, Y. Higashi and M. Ochi (2015). "Mesenchymal-stem-cell-derived exosomes accelerate skeletal muscle regeneration." FEBS Letters 589(11): 1257-1265. 90. Almeria, C., R. Weiss, M. Roy, C. Tripisciano, C. Kasper, V. Weber and D. Egger (2019). "Hypoxia Conditioned Mesenchymal Stem Cell-Derived Extracellular Vesicles Induce Increased Vascular Tube Formation in vitro." Frontiers in Bioengineering and Biotechnology 7. | - |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/90321 | - |
dc.description.abstract | 相對於傳統平面培養的模式,多細胞球體培養有利於細胞與細胞、細胞與胞外基質之間的訊息傳遞;球體培養也能造成球體內部缺氧而促成缺氧因子的表現及反應。本研究旨在探討人類牙髓幹細胞球體培養是否具有增進血管新生的特性。
研究材料為取自齒顎矯正病人拔除之智齒培養的初代牙髓幹細胞,將細胞培養於幾丁聚醣薄膜使之聚集形成多細胞球體,對照組為傳統培養的平面細胞。研究方法包括牙髓幹細胞形態分析、細胞活性測試、缺氧及血管新生因子之蛋白質表現與mRNA表現分析。利用西方墨點法分析蛋白質表現,並以反轉錄聚合酶連鎖反應分析上游基因的調控變化。此外,本研究尚收集牙髓幹細胞球體之條件培養基,並利用人類臍靜脈內皮細胞進行體外血管形成試驗,探究細胞分泌物質是否具有促進血管新生的潛能。 研究結果顯示牙髓幹細胞球體形成初期結構較為鬆散;隨著時間推進,球體尺寸增加,細胞之間的排列也更加緊密。在經過多天培養後,將球體細胞分散後再回種培養,發現仍具細胞活性並能持續增殖。蛋白質分析顯示成球培養模式可誘發牙髓幹細胞處於缺氧環境,成球培養二天後的缺氧誘導因子-1α(HIF-1α)蛋白質表現上升,培養三天後差異仍然顯著;血管內皮生長因子-A (VEGF-A)在成球培養二天後也呈現顯著上升;至於血管生成素1 (Angptn-1)的表現在成球培養下僅有些許上升的趨勢。反轉錄聚合酶連鎖反應分析顯示:成球牙髓幹細胞的缺氧誘導因子1α基因的mRNA在第一天即有顯著上升;血管生成素1基因在第三天的mRNA表現顯著上升;血管生成素2的mRNA表現於第一天顯著提高。其他相關血管新生因子之mRNA表現,包括血管內皮生長因子A與鹼性纖維母細胞生長因子,在平面培養與球體培養的牙髓幹細胞表現皆無顯著差別。在體外血管形成試驗方面的結果,培養成球牙髓幹細胞之條件培養基能夠顯著促進人類臍靜脈內皮細胞形成管狀結構,具有促進內皮細胞血管新生的功能。 總結而言,本研究闡明在幾丁聚醣薄膜上培養之球體牙髓幹細胞的血管新生效應優於傳統平面培養細胞,具有應用於傷口修復以及組織再生的潛能。 | zh_TW |
dc.description.abstract | Three-dimensional (3D) spheroid culture has been recognized to support intercellular communication and mimick native tissue microenvironment in vivo. This study aimed to investigate angiogenic potential of human dental pulp stem cells (DPSCs) cultured as 3D multicellular spheroids.
Primary DPSCs were isolated from dental pulp tissues extracted from orthodontic patients' third molars. DPSCs were cultured on chitosan membranes to form multicellular spheroids, while the control cells were cultured as 2D monolayers on conventional tissue culture polystyrene (TCPS) dishes. The comparison between spheroids and 2D monolayer cultures included cell morphology, cell viability, protein expression and mRNA expression of hypoxia- and angiogenesis-related factors. The conditioned media from the cultured DPSCs were collected for tube formation assay on human umbilical vein endothelial cells (HUVECs) to evaluate the angiogenic potential of secreted factors. The results demonstrated that the initially formed DPSC spheroids exhibited a loose structure. As time progressed, the spheroid size increased, and the arrangement of cells became more compact. The size of individual cells of spheroid culture was smaller than that of 2D monolayer culture. After dissociation and reseeding, the cells from spheroids could proliferate steadily with culture time. The spheroid culture induced a hypoxic environment for DPSCs, as evidenced by an upregulation of hypoxia-inducible factor-1α (HIF-1α) protein expression in the 2-day spheroid culture, and the difference was still significant in the 3-day culture. The protein expression of vascular endothelial growth factor-A (VEGF-A) was also enhanced in 2-day spheroid culture, while the angiopoietin-1 (Angptn-1) exhibited only a slight increasing trend. The reverse transcription polymerase chain reaction (RT-PCR) revealed that the mRNA levels of hypoxia-inducible factor-1α and angiopoietin-2 significantly increased in the 1-day spheroid culture, the angiopoietin-1 gene showed significant expression in the 3-day culture. The mRNA expression of other angiogenesis-related factors, including vascular endothelial growth factor-A and basic fibroblast growth factor, showed no significant differences between monolayer and spheroid DPSC cultures. In terms of the results of the tube formation assay, conditioned media from spheroid-cultured DPSCs significantly promoted the formation of tubular structures by human umbilical vein endothelial cells, indicating a pro-angiogenic function in promoting endothelial cell blood vessel formation. In summary, our study demonstrated that the angiogenic potential of DPSCs cultured as spheroids on chitosan membranes was superior to that of conventional monolayer culture, highlighting their potential in the application for wound healing and tissue regeneration. | en |
dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2023-09-26T16:15:29Z No. of bitstreams: 0 | en |
dc.description.provenance | Made available in DSpace on 2023-09-26T16:15:29Z (GMT). No. of bitstreams: 0 | en |
dc.description.tableofcontents | 誌謝 i
摘要 iii ABSTRACT v 目錄 vii 圖表目錄 xi 第1章 緒論 1 1.1 研究動機 1 1.2 研究背景 1 1.2.1 牙髓幹細胞 1 1.2.2 平面與成球細胞培養 2 1.3 研究目的 2 第2章 文獻回顧 3 2.1 立體多細胞球體 3 2.1.1 立體多細胞球之形成方法 3 2.1.2 立體多細胞球特徵與優勢 3 2.1.3 立體多細胞球之型態分類與影響 4 2.1.4 立體細胞球培養與血管新生 4 2.2 幾丁聚醣用於細胞球體形成 5 2.2.1 利用幾丁聚醣使細胞成球之機制 5 2.2.2 球體間質幹細胞之潛能 6 2.3 血管新生 6 2.3.1 血管新生之生理機制 6 2.3.2 血管新生因子 7 2.3.3 缺氧與血管新生 8 2.4 人體牙髓幹細胞之血管新生潛能 9 2.4.1 牙髓與牙髓幹細胞 9 2.4.2 牙髓幹細胞促進血管新生的機制 9 2.4.3 牙髓幹細胞之血管新生之應用 10 第3章 研究方法 11 3.1 細胞分離與培養 11 3.1.1 細胞球體培養盤製作 11 3.1.2 細胞球體形態分析 11 3.1.3 細胞球體分離之個別細胞形態分析 12 3.1.4 回種細胞之生長測試 12 3.2 西方墨點法 13 3.2.1 樣品收集與定量 13 3.2.2 蛋白質電泳分離、轉漬、封閉、及抗體反應 14 3.2.3 檢測與分析 15 3.3 聚合酶連鎖反應 15 3.3.1 樣品收集 15 3.3.2 核糖核酸萃取 15 3.3.3 反轉錄聚合酶連鎖反應 16 3.3.4 瓊脂凝膠電泳分析 17 3.4 血管形成試驗 17 3.4.1 樣品收集 18 3.4.2 人類臍帶靜脈內皮細胞 18 3.4.3 血管形成試驗培養盤備製 18 3.4.4 血管形成試驗與分析 19 3.5 統計分析 19 第4章 實驗結果 20 4.1 形態分析 20 4.1.1 3D細胞球體形態 20 4.1.2 成球培養對單獨細胞形態的影響 21 4.1.3 回種牙髓幹細胞之生長測試 21 4.2 蛋白質分析 22 4.2.1 HIF-1α之蛋白質表現 22 4.2.2 VEGFA之蛋白質表現 23 4.2.3 Angiopoietin-1之蛋白質表現 23 4.3 反轉錄聚合酶連鎖反應結果 24 4.3.1 HIF1A基因之mRNA表現 24 4.3.2 VEGFA基因之mRNA表現 24 4.3.3 Angiopoietin-1、2基因之mRNA表現 25 4.3.4 Angiogenin之基因表現 25 4.3.5 Basic fibroblast growth factor之基因表現 26 4.4 體外血管形成試驗結果 26 第5章 討論 28 5.1 球體培養對牙髓幹細胞形態之影響 28 5.2 細胞球體缺氧及促血管新生因子基因調控 30 5.3 條件培養基之角色及其促血管新生之效益 34 第6章 結論 36 第7章 未來研究方向 38 第8章 參考文獻 39 圖表目錄 表 1半定量-反轉錄-聚合酶連鎖反應所使用之引子序列 45 圖 1 幾丁聚醣培養之牙髓幹細胞球 46 圖 2 牙髓幹細胞球與平面牙髓幹細胞分離後之細胞形態 47 圖 3 分散的牙髓幹細胞回種於TPCS一天後之影像 48 圖 4回種牙髓幹細胞之生長測試 49 圖 5 缺氧誘導因子1α之蛋白質表現 50 圖 6 血管內皮生長因子之蛋白質表現 51 圖 7 血管生成素1之蛋白質表現 52 圖 8缺氧誘導因子1A之基因表現 53 圖 9 血管內皮生長因子A之基因表現 54 圖 10 血管生成素-1之基因表現 55 圖 11 血管生成素-2之基因表現 56 圖 12 血管生成因子之基因表現 57 圖 13 鹼性纖維母細胞生長因子之基因表現 58 圖 14 ImageJ 分析示意圖 59 圖 15 血管生成試驗量化分析以及形成管道形態的代表性圖示 60 | - |
dc.language.iso | zh_TW | - |
dc.title | 人類牙髓幹細胞球體之體外血管新生效應探討 | zh_TW |
dc.title | The Angiogenic Effects of Human Dental Pulp Stem Cells Spheroids in Vitro | en |
dc.type | Thesis | - |
dc.date.schoolyear | 111-2 | - |
dc.description.degree | 碩士 | - |
dc.contributor.oralexamcommittee | 楊台鴻;鄭景暉 | zh_TW |
dc.contributor.oralexamcommittee | Tai-Horng Young;Jiiang-Huei Jeng | en |
dc.subject.keyword | 幾丁聚醣,牙髓幹細胞,球體培養,體外血管形成試驗, | zh_TW |
dc.subject.keyword | chitosan,dental pulp stem cells (DPSCs),spheroid culture,tube formation assay, | en |
dc.relation.page | 60 | - |
dc.identifier.doi | 10.6342/NTU202301684 | - |
dc.rights.note | 未授權 | - |
dc.date.accepted | 2023-07-18 | - |
dc.contributor.author-college | 醫學院 | - |
dc.contributor.author-dept | 臨床牙醫學研究所 | - |
顯示於系所單位: | 臨床牙醫學研究所 |
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