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研究生:張曉旻
研究生(外文):Chang, Hsiao-Min
論文名稱:利用鼠胚纖維母細胞建立毛囊幹細胞體外自我更新之培養環境
論文名稱(外文):Maintenance of differentiation capability of mouse hair follicle stem cell with embryonic fibroblast feeder
指導教授:沈家寧詹爾昌
指導教授(外文):Shen, Chia-NingChan, Err-Cheng
學位類別:碩士
校院名稱:長庚大學
系所名稱:醫學生物技術研究所
學門:醫藥衛生學門
學類:醫學技術及檢驗學類
論文種類:學術論文
論文出版年:2006
畢業學年度:94
語文別:中文
論文頁數:89
中文關鍵詞:毛囊幹細胞鼠胚纖維母細胞多潛能性
外文關鍵詞:hair follicle stem cellmouse embryonic fibroblastmultipotency
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近幾年研究結果指出毛囊幹細胞可能存在毛囊的髮凸(bulge)結構裏,所以也被命名為髮凸幹細胞(bulge stem cells),經由動物體內的細胞系追蹤(lineage tracing)實驗或皮膚移植實驗結果顯示,毛囊幹細胞具有形成角質細胞以修復受傷表皮、分化成皮脂腺細胞以及再生毛髮的多潛能的分化能力。近期研究也發現毛囊幹細胞可能表現特殊細胞表面標誌如CD34、CD49f等,因此可基於此特性而藉由流式細胞儀從皮膚分離出來,然而並未有研究確切指出毛囊幹細胞可在體外環境(in vitro)中培養增生並可維持其多潛能分化的特性,進一步地並可在植入皮膚組織而後仍可以長出毛髮。因此本論文目標在嘗試建立體外培養環境來提供毛囊幹細胞進行增生及分化,並探討各種調控因子對於毛囊幹細胞在毛囊分化過程中的影響。由目前結果指出,在E13.5與E18.5鼠胚胎纖維母細胞(mouse embryonic fibroblast)的共同培養下可模擬毛囊發育的微環境(microenvironment),而維持成鼠毛囊幹細胞未分化的特性並可在體外環境增生,相反地與初生鼠皮膚纖維母細胞(dermal fibroblasts; DF)共同培養下,成鼠毛囊幹細胞容易分化為角質細胞,顯示不同滋養層細胞可能藉由分泌調控因子的不同而影響幹細胞生長與分化狀態,進ㄧ步地如將培養於E18.5 胚胎纖維母細胞上之毛囊幹細胞注射入初生鼠之上唇皮膚的真皮層中作體外器官培養(ex vivo organ culture),在四星期後,發現會有細毛產生,此結果說明在E18.5 胚胎纖維母細胞的共同培養環境,除能促進毛囊幹細胞增生外,還能維持其分化特性,因此未來可進一步利用這樣的培養系統來探討微環境中的各種因子對於毛囊幹細胞分化及自我更新的調控。
Recent studies using in vivo engraftment studies have revealed that hair follicle stem cells, which reside in the bulge of the hair follicle, could differentiate into multiple cell types including, sebaceous glands, entire hair follicle and epidermis. Moreover, several groups have successfully isolated hair follicle stem cells recently with specific cell surface markers such as CD34 and CD49f. However, there is no evidence showing that isolated hair follicle stem cells could maintain its differentiation capability in vitro. Therefore, in the current studies, , with the utilization of two-step trypsin digestion method and fluorescence-activated cell sorting (FACS)based on the expression of CD34 and CD49f, we have established a method to isolate hair follicle stem cells from neonatal/adult mice.. When coculturing isolated hair follicle stem cells with different feeder cells including mouse embryonic fibroblasts (MEFs) isolated from various embryonice stage , neonatal dermal fibroblasts (DFs), and 3T3 cells, we found that inE13.5 and E18.5 MEF-cocuturing condition, stem cell colonies were formed with appearance of undifferentiated cell morphology. In contrast, when hair follicle stem cells grown onto DF together with EGF and calcium, stem cells differentiated into keratinocyte-like cells. Furthermore, injection of stem cell colonies obtained from E18.5 MEF-co-culturing condition into neonatal upper-lip, , we found that fine hairs were grown from lips after four-week’s culture. In summary, our results suggest that embryonic fibroblast cells can possibly provide a suitable microenvironment to expand and to maintain the multipotency of adult hair follicle stem cells in vitro.
目錄
中文摘要.............................................................1
英文摘要.............................................................3
中英名詞縮寫對照表.....................................................4
第一章 緒論..........................................................5
1.1. 前言.........................................................5
1.2. 毛囊發育與型態發生.............................................6
1.3. 毛髮生長週期..................................................9
1.4. 毛囊形態發生與出生後更新週期中之分子調控機轉.....................11
1.4.1. 真皮乳突....................................................12
1.4.2. Wnt signaling..............................................13
1.4.3. Bone morphogenetic proteins (BMP) signaling................14
1.4.4. Hoxc13.....................................................16
1.5. 毛囊幹細胞...................................................18
1.6. 實驗動機....................................................21
1.7. 實驗目的....................................................23
第二章 實驗材料與方法................................................24
第三章 實驗結果.....................................................33
第四章 討論.........................................................42
參考文獻............................................................50
圖..................................................................59
表..................................................................88
Alonso, L. and E. Fuchs (2003). "Stem cells in the skin: waste not, Wnt not." Genes Dev 17(10): 1189-200.

Andl, T., S. T. Reddy, et al. (2002). "WNT signals are required for the initiation of hair follicle development." Dev Cell 2(5): 643-53.

Alexander Awgulewitsch (2003) Hox in hair growth and development. Naturwissenschaften 90, 193-211

Barrandon, Y. and H. Green (1987). "Three clonal types of keratinocyte with different capacities for multiplication." Proc Natl Acad Sci U S A 84(8): 2302-6.

Blanpain, C., W. E. Lowry, et al. (2004). "Self-renewal, multipotency, and the existence of two cell populations within an epithelial stem cell niche." Cell 118(5): 635-48.

Blessing, M., P. Schirmacher, et al. (1996). "Overexpression of bone morphogenetic protein-6 (BMP-6) in the epidermis of transgenic mice: inhibition or stimulation of proliferation depending on the pattern of transgene expression and formation of psoriatic lesions." J Cell Biol 135(1): 227-39.

Botchkarev, V. A., N. V. Botchkareva, et al. (1999). "Noggin is a mesenchymally derived stimulator of hair-follicle induction." Nat Cell Biol 1(3): 158-64.

Capecchi MR (1997) Hox genes and mammalian development. Cold Spring Harbor Symp Quant Biol 62, 273-281

Chen W, Kelly MA, et al. (1997). "Exocrine gland dysfunction in MC5R-deficient mice: evidence for coordinated regulation of exocrine gland function by melanocortin peptides." Cell 91, 789–98.

Claudia Bagutti, Caroline Hutter, et al. (2001). "Dermal fibroblast-derived growth factors restore the ability of β1 integrin-deficient embryonal stem cells to differentiate into keratinocytes." Developmental Biology 231, 321-333

Cotsarelis, G., T. T. Sun, et al. (1990). "Label-retaining cells reside in the bulge area of pilosebaceous unit: implications for follicular stem cells, hair cycle, and skin carcinogenesis." Cell 61(7): 1329-37.

DasGupta, R. and E. Fuchs (1999). "Multiple roles for activated LEF/TCF transcription complexes during hair follicle development and differentiation." Development 126(20): 4557-68.

Fuchs, E. and D. W. Cleveland (1998). "A structural scaffolding of intermediate filaments in health and disease." Science 279(5350): 514-9.

Gandarillas, A. (2000). "Epidermal differentiation, apoptosis, and senescence: common pathways?" Exp Gerontol 35(1): 53-62.

Godwin AR, Capecchi MR (1998) Hoxc13 mutant mice lack external hair. Genes Dev 12, 11-20

Horace Rhee, Lisa Polak, Elaine Fuchs (2006). " Lhx2 maintains stem cell character in hair follicles. " Science 312(5782): 1946-1949.

Huelsken, J., R. Vogel, et al. (2001). "beta-Catenin controls hair follicle morphogenesis and stem cell differentiation in the skin." Cell 105(4): 533-45.

Jahoda, C. A. and R. F. Oliver (1984). "Vibrissa dermal papilla cell aggregative behaviour in vivo and in vitro." J Embryol Exp Morphol 79: 211-24.

Jahoda, C. A., K. A. Horne, et al. (1984). "Induction of hair growth by implantation of cultured dermal papilla cells." Nature 311(5986): 560-2.

Jahoda, C. A. (1992). "Induction of follicle formation and hair growth by vibrissa dermal papillae implanted into rat ear wounds: vibrissa-type fibres are specified." Development 115(4): 1103-9.

Kalinin, A. E., A. V. Kajava, et al. (2002). "Epithelial barrier function: assembly and structural features of the cornified cell envelope." Bioessays 24(9): 789-800.

Kishimoto, J., R. E. Burgeson, et al. (2000). "Wnt signaling maintains the hair-inducing activity of the dermal papilla." Genes Dev 14(10): 1181-5.

Krumlauf R (1994) Hox genes in vertebrate development. Cell 78, 191-201

Kulessa H, Turk G, Hogan BL (2000) Inhibition of Bmp signaling affects growth and differentiation in the anagen hair follicle. EMBO J 19, 6664-6674

Lichti, U., W. C. Weinberg, et al. (1993). "In vivo regulation of murine hair growth: insights from grafting defined cell populations onto nude mice." J Invest Dermatol 101(1 Suppl): 124S-129S.

Lindner, G., V. A. Botchkarev, et al. (1997). "Analysis of apoptosis during hair follicle regression (catagen)." Am J Pathol 151(6): 1601-17.

Lyons, K. M., R. W. Pelton, et al. (1989). "Patterns of expression of murine Vgr-1 and BMP-2a RNA suggest that transforming growth factor-beta-like genes coordinately regulate aspects of embryonic development." Genes Dev 3(11): 1657-68.

Lyons, K. M., R. W. Pelton, et al. (1990). "Organogenesis and pattern formation in the mouse: RNA distribution patterns suggest a role for bone morphogenetic protein-2A (BMP-2A)." Development 109(4): 833-44.

Maretto, S., M. Cordenonsi, et al. (2003). "Mapping Wnt/beta-catenin signaling during mouse development and in colorectal tumors." Proc Natl Acad Sci U S A 100(6): 3299-304.

Mark M. Rijli FM, Chambon P (1997) Homeobox genes in embryogenesis and pathogenesis. Pediatr Res 42, 421-429

Martin, P. (1997). "Wound healing – aiming for perfect skin regeneration. " Science 276, 75-81

Massague, J. and Y. G. Chen (2000). "Controlling TGF-beta signaling." Genes Dev 14(6): 627-44.

Matsuzaki, T. and K. Yoshizato (1998). "Role of hair papilla cells on induction and regeneration processes of hair follicles." Wound Repair Regen 6(6): 524-30.

McGinnis W, Krumlauf R (1992) Homeobox genes and axial patterning. Cell 68,283-302

Morris, R. J. and C. S. Potten (1994). "Slowly cycling (label-retaining) epidermal cells behave like clonogenic stem cells in vitro." Cell Prolif 27(5): 279-89.

Morris, R. J., Y. Liu, et al. (2004). "Capturing and profiling adult hair follicle stem cells." Nat Biotechnol 22(4): 411-7.

Nohe, A., S. Hassel, et al. (2002). "The mode of bone morphogenetic protein (BMP) receptor oligomerization determines different BMP-2 signaling pathways." J Biol Chem 277(7): 5330-8.

Nohe, A., E. Keating, et al. (2004). "Signal transduction of bone morphogenetic protein receptors." Cell Signal 16(3): 291-9.

O'Shaughnessy, R. F., W. Yeo, et al. (2004). "The WNT signalling modulator, Wise, is expressed in an interaction-dependent manner during hair-follicle cycling." J Invest Dermatol 123(4): 613-21.

Oshima, H., A. Rochat, et al. (2001). "Morphogenesis and renewal of hair follicles from adult multipotent stem cells." Cell 104(2): 233-45.Potten, C. S. (1975). "Epidermal transit times." Br J Dermatol 93(6): 649-58.

Paus, R., S. Muller-Rover, et al. (1999). "A comprehensive guide for the recognition and classification of distinct stages of hair follicle morphogenesis." J Invest Dermatol 113(4): 523-32.

Paus, R., N. Krejci-Papa, et al. (1994). "Correlation of proteolytic activities of organ cultured intact mouse skin with defined hair cycle stages." J Dermatol Sci 7(3): 202-9.

R Barthel, D Aberdam. (2005). "Epidermal stem cells" JEADV 19: 405-413

Reddi, A. H. (1998). Role of morphogenetic proteins in skeletal tissue engineering and regeneration." Nat Biotechnol 16(3): 247-52.

Rheinwald, J. G. and H. Green (1975). "Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells." Cell 6(3): 331-43.

Rheinwald, J. G. and H. Green (1977). "Epidermal growth factor and the multiplication of cultured human epidermal keratinocytes." Nature 265(5593): 421-4.

Scott MP (1992) Vertebrate homeobox gene nomenclature. Cell 71, 551-553

Schmidt-Ullrich, R. and R. Paus (2005). "Molecular principles of hair follicle induction and morphogenesis." Bioessays 27(3): 247-61.

Seiberg, M., J. Marthinuss, et al. (1995). "Changes in expression of apoptosis-associated genes in skin mark early catagen." J Invest Dermatol 104(1): 78-82.

Stenn, K. S. and R. Paus (2001). "Controls of hair follicle cycling." Physiol Rev 81(1): 449-494.

St-Jacques, B., H. R. Dassule, et al. (1998). "Sonic hedgehog signaling is essential for hair development." Curr Biol 8(19): 1058-68.

Takahashi, H. and T. Ikeda (1996). "Transcripts for two members of the transforming growth factor-beta superfamily BMP-3 and BMP-7 are expressed in developing rat embryos." Dev Dyn 207(4): 439-49.

Taylor, G., M. S. Lehrer, et al. (2000). "Involvement of follicular stem cells in forming not only the follicle but also the epidermis." Cell 102(4): 451-61.

Tkatchenko AV, Visconti RP, et al. (2001) "Overexpression of Hoxc13 in differentiating keratinocytes results in downregulation of a novel hair keratin gene cluster and alopecia." Development 128, 1547-1558

Trempus CS, Morris RJ, et al. (2003) "Enrichment for living murine keratinocytes from the hair follicle bulge with the cell surface marker CD34." J Invest Dermatol. 120(4):501-11.

Tumbar, T., G. Guasch, et al. (2004). "Defining the epithelial stem cell niche in skin." Science 303(5656): 359-63.

Van Genderen, C., R. M. Okamura, et al. (1994). "Development of several organs that require inductive epithelial-mesenchymal interactions is impaired in LEF-1-deficient mice." Genes Dev 8(22): 2691-703.

Wall, N. A., M. Blessing, et al. (1993). "Biosynthesis and in vivo localization of the decapentaplegic-Vg-related protein, DVR-6 (bone morphogenetic protein-6)." J Cell Biol 120(2): 493-502.

Watt, F. M. (1998). "Epidermal stem cells: markers, patterning and the control of stem cell fate." Philos Trans R Soc Lond B Biol Sci 353(1370): 831-7.

Wilson, P. A. and A. Hemmati-Brivanlou (1995). "Induction of epidermis and inhibition of neural fate by Bmp-4." Nature 376(6538): 331-3.

Wodarz, A. and R. Nusse (1998). "Mechanisms of Wnt signaling in development." Annu Rev Cell Dev Biol 14: 59-88.

Zheng Y, Prouty SM, et al. (2001). "Scd3--a novel gene of the stearoyl-CoA desaturase family with restricted expression in skin." Genomics 71(2):182-91.
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