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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/22967Full metadata record
| ???org.dspace.app.webui.jsptag.ItemTag.dcfield??? | Value | Language |
|---|---|---|
| dc.contributor.advisor | 謝銘鈞(Ming-Jium Shieh) | |
| dc.contributor.author | Hau-Ming Jan | en |
| dc.contributor.author | 詹皓名 | zh_TW |
| dc.date.accessioned | 2021-06-08T04:35:38Z | - |
| dc.date.copyright | 2009-08-20 | |
| dc.date.issued | 2009 | |
| dc.date.submitted | 2009-08-18 | |
| dc.identifier.citation | 1. Hwang J, Mehrani T, Millar SE, Morasso MI: Dlx3 is a crucial regulator of hair follicle differentiation and cycling. Development 2008, 135(18):3149-3159.
2. Ito M, Yang Z, Andl T, Cui C, Kim N, Millar SE, Cotsarelis G: Wnt-dependent de novo hair follicle regeneration in adult mouse skin after wounding. Nature 2007, 447(7142):316-320. 3. Choi J, Southworth LK, Sarin KY, Venteicher AS, Ma W, Chang W, Cheung P, Jun S, Artandi MK, Shah N et al: TERT promotes epithelial proliferation through transcriptional control of a Myc- and Wnt-related developmental program. PLoS Genet 2008, 4(1):e10. 4. Sarin KY, Cheung P, Gilison D, Lee E, Tennen RI, Wang E, Artandi MK, Oro AE, Artandi SE: Conditional telomerase induction causes proliferation of hair follicle stem cells. Nature 2005, 436(7053):1048-1052. 5. Gordon JW, Ruddle FH: GENE-TRANSFER INTO MOUSE EMBRYOS - PRODUCTION OF TRANSGENIC MICE BY PRONUCLEAR INJECTION. Methods in Enzymology 1983, 101:411-433. 6. Palmiter RD, Wilkie TM, Chen HY, Brinster RL: TRANSMISSION DISTORTION AND MOSAICISM IN AN UNUSUAL TRANSGENIC MOUSE PEDIGREE. Cell 1984, 36(4):869-877. 7. Rusconi S, Kohler G: TRANSMISSION AND EXPRESSION OF A SPECIFIC PAIR OF REARRANGED IMMUNOGLOBULIN MU-GENES AND KAPPA-GENES IN A TRANSGENIC MOUSE LINE. Nature 1985, 314(6009):330-334. 8. Cheng H, Zhu JL, Zeng X, Jing Y, Zhang XZ, Zhuo RX: Targeted Gene Delivery Mediated by Folate-polyethylenimine-block-poly(ethylene glycol) with Receptor Selectivity. Bioconjug Chem 2009. 9. Shim MS, Kwon YJ: Acid-Responsive Linear Polyethylenimine for Efficient, Specific, and Biocompatible siRNA Delivery. Bioconjug Chem 2009. 10. Zhao QQ, Chen JL, Lv TF, He CX, Tang GP, Liang WQ, Tabata Y, Gao JQ: N/P ratio significantly influences the transfection efficiency and cytotoxicity of a polyethylenimine/chitosan/DNA complex. Biol Pharm Bull 2009, 32(4):706-710. 11. Domashenko A, Gupta S, Cotsarelis G: Efficient delivery of transgenes to human hair follicle progenitor cells using topical lipoplex. Nat Biotechnol 2000, 18(4):420-423. 12. Itaka K, Kataoka K: Recent development of nonviral gene delivery systems with virus-like structures and mechanisms. Eur J Pharm Biopharm 2009, 71(3):475-483. 13. Ko YT, Bhattacharya R, Bickel U: Liposome encapsulated polyethylenimine/ODN polyplexes for brain targeting. J Control Release 2009, 133(3):230-237. 14. Schlake T: Determination of hair structure and shape. Semin Cell Dev Biol 2007, 18(2):267-273. 15. Trempus CS, Morris RJ, Bortner CD, Cotsarelis G, Faircloth RS, Reece JM, Tennant RW: Enrichment for living murine keratinocytes from the hair follicle bulge with the cell surface marker CD34. J Invest Dermatol 2003, 120(4):501-511. 16. Morris RJ, Liu Y, Marles L, Yang Z, Trempus C, Li S, Lin JS, Sawicki JA, Cotsarelis G: Capturing and profiling adult hair follicle stem cells. Nat Biotechnol 2004, 22(4):411-417. 17. Wong K, Sun G, Zhang X, Dai H, Liu Y, He C, Leong KW: PEI-g-chitosan, a novel gene delivery system with transfection efficiency comparable to polyethylenimine in vitro and after liver administration in vivo. Bioconjug Chem 2006, 17(1):152-158. 18. Ryser HJ: A membrane effect of basic polymers dependent on molecular size. Nature 1967, 215(5104):934-936. 19. Funhoff AM, van Nostrum CF, Koning GA, Schuurmans-Nieuwenbroek NM, Crommelin DJ, Hennink WE: Endosomal escape of polymeric gene delivery complexes is not always enhanced by polymers buffering at low pH. Biomacromolecules 2004, 5(1):32-39. 20. Schlake T, Sick S: Canonical WNT signalling controls hair follicle spacing. Cell Adh Migr 2007, 1(3):149-151. 21. Ouji Y, Yoshikawa M, Moriya K, Ishizaka S: Effects of Wnt-10b on hair shaft growth in hair follicle cultures. Biochem Biophys Res Commun 2007, 359(3):516-522. 22. Sick S, Reinker S, Timmer J, Schlake T: WNT and DKK determine hair follicle spacing through a reaction-diffusion mechanism. Science 2006, 314(5804):1447-1450. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/22967 | - |
| dc.description.abstract | 基因轉移到毛囊是一種有潛力的技術來治療皮膚疾病。因為皮膚有非常低的滲透性，所以在活體的皮膚毛囊細胞上，非病毒基因轉殖才被認為是一種困難的方法。在此，我們利用聚乙烯亞胺 (PEI)來包覆人類的端粒酶逆轉錄酶基因 (hTERT)，然後利用塗抹在皮膚表面的方式，可以成功的將hTERT這個基因送到正常大鼠皮膚組織和受傷過的皮膚組織的毛囊細胞，讓這些細胞表現hTERT這個基因。我們發現，在正常皮膚組織的毛囊中，hTERT的表達會使毛囊幹細胞增生，進而加速毛髮的生長。另一方面，在受傷的皮膚組織的毛囊中，hTERT的表達則是會造成毛囊的再生。本研究利用創新的基因轉移的方法，並提供了一種實際的解決方法治療皮膚疾病。 | zh_TW |
| dc.description.abstract | Gene transfer to hair follicles is a potential technology to treat skin disease. Non-viral transfection is considered a difficult method to deliver genes into hair follicles in vivo because of the extremely low permeability of skin. We utilized polyethylenimine (PEI) to transfect the human telomerase reverse transcriptase (hTERT) in hair follicle cells in normal and wounding rat skin tissues. We found that expression of hTERT in normal skin tissue could increase the number of proliferating stem cells in hair follicles. On the other hand, expression of hTERT in wounding skin tissue can promote hair follicle de novo formation. These studies might not only reveal an innovative method for gene transfer, but also provide a practical solution for skin disease therapy. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T04:35:38Z (GMT). No. of bitstreams: 1 ntu-98-R96548012-1.pdf: 4109469 bytes, checksum: f62c7421366ca54251623866244f459e (MD5) Previous issue date: 2009 | en |
| dc.description.tableofcontents | 目 錄
口試委員會審定書………………………………………………………………… i 誌謝………………………………………………………………………………….. ii 中文摘要……………………………………………………………………….. iii 英文摘要…………………………………………………………………………….. iv 1. Introduction：…………………………………………………………………. 1~2 2. Material and method：…………………………………………..…………. 3~13 2.1 Antibodies and Chemicals. ………………………………………………. 3~4 2.2 Magnetic Labeling and Separation of Cell………………………………. 4∼5 2.3 Flow cytometry…………………………………………………………... 5~6 2.4 Cell culture of hair follicle stem cell…………………………………… 6~8 2.5 Immunofluorescent staining ………………………...………………… 8~9 2.6 Colony formation…………………………………………………………... 9 2.7 Transfection in vitro……………………………………………………… 9~10 2.8 Cytotoxicity - MTT assay………………………………………………. 10~11 2.9 Histochemical assay of ß-galactosidase activity…………………………… 11 2.10 In vivo mouse hair follicle transfection………………………………………. 11 2.11 Immunohistochemistry staining and H&E staining……………………… 12 2.12 Counting of hair length……………………………………………………12~13 2.13 Whole-mount hair follicle neogenesis assay. ………………………… 13 3. Result……………………………………………………………………… 13~22 3.1 Purification of hair follicle stem cells…………………………………… 13~15 3.2 In vitro transfection efficiency and cytotoxicity of PEI…………………..15~16 3.3 The function of hTERT in vitro in rat keratinocyte……………………… 17~18 3.4 In vivo transfection efficiency of PEI………………………… 18~19 3.5 The function of hTERT in vivo in rat skin tissue…………….…………...… 19~21 3.6 hTERT promote de novo follicle formation. ………...…………21~22 4. Discussions……………………………………………………………… 22~25 4.1 PEI serves as a promising vector for gene therapy in skin disease……… 22~23 4.2 TERT serve as a hair growth enhancer…………………………………..23~24 4.3 TERT and Wnt-signaling pathway………………………………24~25 5. Conclusion …………………………………………………………………25 6. Reference ……………………………………………………………… 26~27 附錄……………………………………………………………………….….…. 28~42 圖目錄 Figure 1. Hair follicle stem cells were sorted by MACS………….………28 Figure 2. The expression of integrin α6…………………………...…………………29 Figure 3. The expression of integrin ß1……………………………………………30 Figure 4. The expression of CD34…………………………………………………38 Figure 5. Clonal growth assays of keratinocytes…………………………………….39 Figure 6. EGFP reported gene expression in hair follicle stem cells…………….…40 Figure 7. hTERT reported gene expression in hair follicle stem cells………....41~42 Figure 8. Cytotoxicity test and transfection efficiency in hair follicle stem cells.…43. Figure 9. In vivo transfection of rat hair follicles…………………………………….44 Figure 10. H&E stain and length of hair after in vivo transfection………………….45 Figure 11 Immunofluorescent staining for PCNA………………………………….46 Figure 12. Expression of hTERT potentiates de novo follicle formation at day10......47 Figure 13. Expression of hTERT potentiates de novo follicle formation at day 40…………………………………………………………………………………48~49 | |
| dc.language.iso | en | |
| dc.subject | 聚乙烯亞胺 | zh_TW |
| dc.subject | 基因轉殖 | zh_TW |
| dc.subject | 人類的端粒酶 | zh_TW |
| dc.subject | 逆轉錄酶 | zh_TW |
| dc.subject | 基因 | zh_TW |
| dc.subject | 毛囊幹細胞 | zh_TW |
| dc.subject | 毛髮生長 | zh_TW |
| dc.subject | hTERT (human telomerase reverse transcriptase) | en |
| dc.subject | hair follicle stem cell | en |
| dc.subject | hair growth | en |
| dc.subject | PEI (polyethylenimine)gene transfer | en |
| dc.title | 高效率基因轉殖技術於毛髮生長上的應用 | zh_TW |
| dc.title | Application of controllable gene expression system in hair growth | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 97-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 賴秉杉(Ping-Shan Lai),婁培人(Pei-Jen Lou),林頌然(Sung-Jan Lin) | |
| dc.subject.keyword | 聚乙烯亞胺,基因轉殖,人類的端粒酶,逆轉錄酶,基因,毛囊幹細胞,毛髮生長, | zh_TW |
| dc.subject.keyword | PEI (polyethylenimine)gene transfer,hTERT (human telomerase reverse transcriptase),hair follicle stem cell,hair growth, | en |
| dc.relation.page | 43 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2009-08-18 | |
| dc.contributor.author-college | 工學院 | zh_TW |
| dc.contributor.author-dept | 醫學工程學研究所 | zh_TW |
| Appears in Collections: | 醫學工程學研究所 | |
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| ntu-98-1.pdf Restricted Access | 4.01 MB | Adobe PDF |
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