請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/33353
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 錢宗良(Chung-Liang Chien) | |
dc.contributor.author | Tsung-Yi Lin | en |
dc.contributor.author | 林宗逸 | zh_TW |
dc.date.accessioned | 2021-06-13T04:36:06Z | - |
dc.date.available | 2006-08-31 | |
dc.date.copyright | 2006-08-31 | |
dc.date.issued | 2006 | |
dc.date.submitted | 2006-07-19 | |
dc.identifier.citation | Barde Y. A., Edgar D., Thoenen H. (1982) Purification of a new neurotrophic factor from mammalian brain. EMBO J 1, 549–553.
Benraiss A., Chmielnicki E., Lerner K., Roh D., Goldman S.A. (2001) Adenoviral brain-derived neurotrophic factor induces both neostriatal and olfactory neuronal recruitment from endogenous progenitor cells in the adult forebrain. J Neurosci 21, 6718–6731. Bibel M., Barde Y. A. (2000) Neurotrophins: key regulators of cell fate and cell shape in the vertebrate nervous system. Genes Dev 14, 2919-2937. Binder D. K., Scharfman H. E. (2004) Brain-derived neurotrophic factor. Growth Factors 22, 123-131. Blum R., Konnerth A. (2005) Neurotrophin-mediated rapid signaling in the central nervous system: mechanisms and functions. Physiology (Bethesda) 20, 70-78. Bullo M., Peeraully M. R., Trayhurn P. (2005) Stimulation of NGF expression and secretion in 3T3-L1 adipocytes by prostaglandins PGD2, PGJ2, and Delta12-PGJ2. Am J Physiol Endocrinol Metab 289, E62-67. Chmielnicki E., Benraiss A., Economides A. N., Goldman S. A. (2004) Adenovirally expressed noggin and brain derived neurotrophic factor cooperate to induce new medium spiny neurons from resident progenitor cells in the adult striatal ventricular zone. J Neurosci 24, 2133–2142. Dechant G., Barde Y. A. (2002) The neurotrophin receptor p75 (NTR): novel functions and implications for diseases of the nervous system. Nat Neurosci 5, 1131–1136. Ernfors P., Ebendal T., Olson L., Mouton P., Stromberg I., Persson H. (1989) A cell line producing recombinant nerve growth factor evokes growth responses in intrinsic and grafted central cholinergic neurons. Proc Natl Acad Sci U S A 86, 4756-4760. Gage F. H., Kawaja M. D., Fisher L. J. (1991) Genetically modified cells: applications for intracerebral grafting. Trends Neurosci 14, 328-333. Gage F. H. (1998) Cell therapy. Nature 392(6679 Suppl), 18-24. Gash D. M., Notter M. F., Okawara S. H., Kraus A. L., Joynt R. J. (1986) Amitotic neuroblastoma cells used for neural implants in monkeys. Science 233, 1420-1422. Green H., Meuth M. (1974) An established pre-adipose cell line and its differentiation in culture. Cell 3, 127-133. Grider M. H., Mamounas L. A., Le W., Shine H. D. (2005) In situ expression of brain-derived neurotrophic factor or neurotrophin-3 promotes sprouting of cortical serotonergic axons following a neurotoxic lesion. J Neurosci Res 82, 404-412. Horellou P., Brundin P., Kaltn P., Mallet J., Bjiirklund A. (1990) In vivo release of DOPA and dopamine from genetically engineered cells grafted to the denervated rat striatum. Neuron 5, 393-402. Hory-Lee F., Russell M., Lindsay R. M., Frank E. (1993) Neurotrophin 3 supports the survival of developing muscle sensory neurons in culture. Proc Natl Acad Sci U S A 90, 2613-2617. Huang E. J., Reichardt L. F. (2001) Neurotrophins: roles in neuronal development and function. Annu Rev Neurosci 24, 677–736. Jainchill J. L., Aaronson S. A., Todaro G. J. (1969) Murine sarcoma and leukemia viruses: assay using clonal lines of contact-inhibited mouse cells. J Virol 4, 549-553. Jerregard H., Akerud P., Arenas E., Hildebrand C. (2000) Fibroblast-like cells from rat plantar skin and neurotrophin-transfected 3T3 fibroblasts influence neurite growth from rat sensory neurons in vitro. J Neurocytol 29, 653-663. Jin Y., Fischer I., Tessler A., Houle J. D. (2002) Transplants of fibroblasts genetically modified to express BDNF promote axonal regeneration from supraspinal neurons following chronic spinal cord injury. Exp Neurol 177, 265-275. Kawaja M. D., Rosenberg M. B., Yoshida K., Gage F. H. (1992) Somatic gene transfer of nerve growth factor promotes the survival of axotomized septal neurons and the regeneration of their axons in adult rats. J Neurosci 12, 2849-2864. Kurozumi K., Nakamura K., Tamiya T., Kawano Y., Kobune M., Hirai S., Uchida H., Sasaki K., Ito Y., Kato K., Honmou O., Houkin K., Date I., Hamada H. (2004) BDNF gene-modified mesenchymal stem cells promote functional recovery and reduce infarct size in the rat middle cerebral artery occlusion model. Mol Ther 9, 189-197. Kurozumi K., Nakamura K., Tamiya T., Kawano Y., Ishii K., Kobune M., Hirai S., Uchida H., Sasaki K., Ito Y., Kato K., Honmou O., Houkin K., Date I., Hamada H. (2005) Mesenchymal stem cells that produce neurotrophic factors reduce ischemic damage in the rat middle cerebral artery occlusion model. Mol Ther 11, 96-104. Lewin G. R., Barde Y. A. (1996) Physiology of the neurotrophins. Ann Rev Neurosci 19, 289–317. Liu Y., Himes B. T., Murray M., Tessler A., Fischer I. (2002) Grafts of BDNF-producing fibroblasts rescue axotomized rubrospinal neurons and prevent their atrophy. Exp Neurol 178, 150-164. McKay R. (1997) Stem cells in the central nervous system. Science 276, 66-71. Momma S., Johansson C. B., and Frisen J. (2000) Get to know your stem cells. Curr Opin Neurobiol 10, 45-49. Nomura T., Honmou O., Harada K., Houkin K., Hamada H., Kocsis J. D. (2005) I.V. infusion of brain-derived neurotrophic factor gene-modified human mesenchymal stem cells protects against injury in a cerebral ischemia model in adult rat. Neuroscience 136, 161-169. Patapoutian A., Reichardt L. F. (2001) Trk receptors: mediators of neurotrophin action. Curr Opin Neurobiol 11, 272-280. Peeraully M. R., Jenkins J. R., Trayhurn P. (2004) NGF gene expression and secretion in white adipose tissue: regulation in 3T3-L1 adipocytes by hormones and inflammatory cytokines. Am J Physiol Endocrinol Metab 287, E331-339. Pencea V., Bingaman K. D., Wiegand S. J., Luskin M. B. (2001) Infusion of brain-derived neurotrophic factor into the lateral ventricle of the adult rat leads to new neurons in the parenchyma of the striatum, septum, thalamus, and hypothalamus. J Neurosci 21, 6706–6717. Radziejewski C., Robinson R. C., DiStefano P. S., Taylor J. W. (1992) Dimeric structure and conformational stability of brain-derived neurotrophic factor and neurotrophin-3. Biochemistry 31, 4431-4436. Rakic P. (1995) Radial versus tangential migration of neuronal clones in the developing cerebral cortex. Proc Natl Acad Sci U S A 92, 11323–11327. Rossner S., Yu J., Pizzo D., Werrbach-Perez K., Schliebs R., Bigl V., Perez-Polo J. R. (1996) Effects of intraventricular transplantation of NGF-secreting cells on cholinergic basal forebrain neurons after partial immunolesion. J Neurosci Res 45, 40-56. Schabitz W. R, Schwab S., Spranger M., Hacke W. (1997) Intraventricular brain-derived neurotrophic factor reduces infarct size after focal cerebral ischemia in rats. J Cereb Blood Flow Metab 17, 500–506. Schabitz W. R., Sommer C., Zoder W., Kiessling M., Schwaninger M., Schwab S. (2000) Intravenous brain-derived neurotrophic factor reduces infarct size and counterregulates Bax and Bcl-2 expression after temporary focal cerebral ischemia. Stroke 31, 2212–2217. Tapia-Arancibia L., Rage F., Givalois L., Arancibia S. (2004) Physiology of BDNF: focus on hypothalamic function. Front Neuroendocrinol 25, 77-107. Taylor G., Lehrer M. S., Jensen P. J., Sun T. T., Lavker R. M. (2000) Involvement of follicular stem cells in forming not only the follicle but also the epidermis. Cell 102, 451-461. Tobias C. A., Dhoot N. O., Wheatley M. A., Tessler A., Murray M., Fischer I. (2001) Grafting of encapsulated BDNF-producing fibroblasts into the injured spinal cord without immune suppression in adult rats. J Neurotrauma 18, 287-301. Tobias C. A., Han S. S., Shumsky J. S., Kim D., Tumolo M., Dhoot N. O., Wheatley M. A., Fischer I., Tessler A., Murray M. (2005) Alginate encapsulated BDNF-producing fibroblast grafts permit recovery of function after spinal cord injury in the absence of immune suppression. J Neurotrauma 22, 138-156. Todaro G. J., Green H. (1963) Quantitative studies of the growth of mouse embryo cells in culture and their development into established lines. J Cell Biol 17, 299-313. Todaro G. J., Green H., Goldberg B. D. (1964) Transformation of properties of an established cell line by SV40 and polyoma virus. Proc Natl Acad Sci U S A 51, 66-73. Westlund K. N., Lu Y., Kadekaro M., Harmann P., Terrell M. L., Pizzo D. P., Hulsebosch C. E., Eisenberg H. M., Perez-Polo J. R. (1995) NGF-producing transfected 3T3 cells: behavioral and histological assessment of transplants in nigral lesioned rats. J Neurosci Res 41, 367-373. Yamashita K., Wiessner C., Lindholm D., Thoenen H., Hossmann K.A. (1997) Post-occlusion treatment with BDNF reduces infarct size in a model of permanent occlusion of the middle cerebral artery in rat. Metab Brain Dis 12, 271-280. Zigova T., Pencea V., Wiegand S. J., Luskin M. B. (1998) Intraventricular administration of BDNF increases the number of newly generated neurons in the adult olfactory bulb. Mol Cell Neurosci 11, 234–245. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/33353 | - |
dc.description.abstract | 腦衍生神經滋養因子(BDNF)在中樞神經系統扮演許多重要的角色。最近有研究發現,BDNF具有誘導腦部成體幹細胞分化的功能,且被認為具有治療腦部損害的潛力。目前,由外加小型滲透幫浦長期導入腦中的BDNF仍有許多待解決的問題。但這些問題可選擇移植能夠持續表達BDNF的細胞而獲得解決之道。
在本研究中,我們同時轉殖小鼠BDNF和綠色螢光蛋白(EGFP)的互補核酸序列(cDNA)進入3T3纖維母細胞株中。在經過neomycin的相似藥物G418的篩選之後,我們建立了兩株穩定獨立表達BDNF和EGFP的細胞株,並命名為3T3-BDNF-EGFP細胞。本實驗中利用免疫染色、西方墨點法和酵素連結抗體吸附法(ELISA)對3T3-BDNF-EGFP細胞進行基因產物分析。另外也透過分析雞胚背根神經結(dorsal root ganglia, DRG)的存活率,來測試由3T3-BDNF-EGFP細胞所分泌的BDNF之生物活性。 從我們得到的免疫染色以及西方墨點法的結果發現,BDNF在3T3-BDNF-EGFP細胞的表達量比控制組來得多。在ELISA的實驗中,也可以發現由3T3-BDNF-EGFP細胞所分泌出BDNF的量具有統計上顯著的增加。而我們進一步利用雞胚的DRG神經元測試BDNF的生物活性,發現由這些3T3-BDNF-EGFP細胞分泌的BDNF能使DRG神經元擁有較高的存活率。 由本研究的結果,我們可以推論3T3-BDNF-EGFP細胞可以穩定表達具有功能性的BDNF。我們認為這些大量表達BDNF,且具有EGFP螢光的細胞,將可以應用在細胞移植治療腦部損傷的動物模式中。 | zh_TW |
dc.description.abstract | Brain-derived neurotrophic factor (BDNF) influences almost all aspects in central nervous system (CNS). Recently, BDNF was discovered as an inducer for adult stem cells and considered as a potential therapeutic agent for the brain injury. However, there are several drawbacks about using injections or minipumps for long-term BDNF treatment. Alternatively, cell mediated BDNF delivery could be one of solutions for it.
In this study, we transfected cDNAs of mouse BDNF and enhanced green fluorescent protein (EGFP) into Swiss albino mouse 3T3 cell line. After selection of neomycin analogue G418, two stable 3T3-BDNF-EGFP cell clones constitutively expressing BDNF and EGFP independently were established. Stable 3T3-BDNF-EGFP cell clones were analyzed by immunocytochemistry, Western blot, and Enzyme-Linked Immunosorbent Assay (ELISA). Besides, the functional test of secreted BDNF activity was also assayed via the viability of chicken DRG neurons. Immunostaining patterns and Western blots of 3T3-BDNF-EGFP cells that we obtained showed more BDNF products than controls. In the ELISA study, the amount of BDNF secreted from 3T3-BDNF-EGFP cells (339 ± 50 pg/ml) was also showed significantly larger than others. Furthermore, we found the higher survival rate in the functional assay of embryonic chicken DRG neurons with BDNF secreted from 3T3-BDNF-EGFP cells. From the present study, we conclude that the stable 3T3-BDNF-EGFP cells could constitutively express functional BDNF in vitro. These BDNF-enriched cells with strong green fluorescence will be useful for further studies, such as the intracerebral cell grafting for brain injuries. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T04:36:06Z (GMT). No. of bitstreams: 1 ntu-95-R93446011-1.pdf: 1337489 bytes, checksum: 61fc1061d8a029b4b1b64f864dc737a4 (MD5) Previous issue date: 2006 | en |
dc.description.tableofcontents | 壹、中英文摘要 ----------------------------------- 1
貳、緒論 ----------------------------------------- 4 參、實驗材料與方法 ------------------------------- 8 肆、實驗結果 ------------------------------------ 15 伍、討論 ---------------------------------------- 19 陸、參考文獻 ------------------------------------ 24 柒、圖表說明 ------------------------------------ 29 捌、附件 ---------------------------------------- 42 | |
dc.language.iso | en | |
dc.title | 建立及特徵分析持續表達腦部神經滋養因子的3T3纖維母細胞株 | zh_TW |
dc.title | Establishment and characterization of constitutively expressed BDNF in genetically modified 3T3 cell line | en |
dc.type | Thesis | |
dc.date.schoolyear | 94-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 歐陽品(Pin Ouyang),盧國賢(Kuo-Shyan Lu) | |
dc.subject.keyword | 腦部神經滋養因子,3T3纖維母細胞株,細胞治療, | zh_TW |
dc.subject.keyword | BDNF,3T3 cell line,cell therapy, | en |
dc.relation.page | 55 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2006-07-20 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 解剖學研究所 | zh_TW |
顯示於系所單位: | 解剖學暨細胞生物學科所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-95-1.pdf 目前未授權公開取用 | 1.31 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。