绒山羊毛囊干细胞的分离培养及诱导分化鉴定

Abstract

Abstract: The aims of this paper were to explore the optimal method of isolating,purifying,and proliferating the Cashmere goat hair follicle stem cells,and their multiple differentiation potentiality. It separated and cultivated the Cashmere goat hair follicle stem cells by two methods,one was the two-step enzyme digestion,another was differential adhesion. Growth curves and colony forming efficiency of passaged the Cashmere goat hair follicle stem cells were determined. The P5 hair follicle stem cells were cultured in vitro under adipogenic and osteogenic inductive environment and induced into adipocytes,osteoblates. The results showed that the isolated hair follicle stem cells were uniform,volume small,paving stone-like,and active proliferative capacity and had S shape of growth curve. Their adipogenic and osteogenic differentiation properties were confirmed by histological staining such as oil red O and alizarim red. Oil red O and alizarim red staining of adipogenic and osteogenic induction group was positive,whereas,control groups were negative.

Key words: Cashmere goat; hair follicle stem cells; induced differentiation

CLC Number:

S813.1⁺1

WANG Hong, GAO Feng, LIU Ying-chun, ZHOU Huan-min. Isolation and Cultures of Hair Follicle Stem Cell from Cashmere Goat and Identification of Induced Differentiation[J]. , 2014, 41(10): 139-144.

References

1 韦林盖,刘宗正,高晋芳,等. 蒙古羊骨髓间充质干细胞的分离培养及多向诱导分化[J]. 中国畜牧兽医,2013,40(8):7~12.
2 史明艳,杨学义,窦忠英. 山羊毛囊干细胞分离培养方法研究[J]. 畜牧兽医学报,2006,37(5):436~440.
3 张群,丛笑倩,张文杰,等. 利用中性蛋白酶消化法扩增毛囊干细胞的实验研究[J]. 上海交通大学学报(医学版),2007,27(4):387~391.
4 谭挺,胡志奇,周洪军. 显微分离培养与免疫磁珠法分离纯化人毛囊干细胞[J]. 中国修复重建外科杂志,2008,22(2):202~205.
5 Amoh Y,Li L,Campillo R,et al. Implanted hair follicle stemcells form Schwann cells that support repair of severed peripheral nerves[J]. Proc Nati Acad Sci USA,2005a,102(49):17734~17738.
6 Amoh Y,Li L,Katsuoka K,et al. Multipotentnestin positive,keratin-negative hair-follicle bulge stem cells can form neurons[J]. Proc Nati Acad Sci USA,2005b,102(15):5530~5534.
7 Aubin-Houzelstein G,Djian-Zaouche J,Bernex F,et al. Melanoblasts' proper location and timed differentiation depend on Notch/RBP-J signaling in postnatal hair follicles[J]. J Invest Dermatol,2008,128(11):2686~2695.
8 Drewa T,Joachimiak R,Bajek A,et al. Hair follicle stem cells can be driven into a urothelial-like phenotype:An experimental study[J]. International Journal of Urology,2013,20(5):537~542.
9 Fuchs E,Tumbar T,Guasch G,et al. Socializing with their neigbors:Stem cells and their niche[J]. Cell,2004,116:769~778.
10 Hilmi A B M,Halim A S,Noor N M,et al. A simple culture method for epithelial stem cells derived from human hair follicle[J]. Cent Eur J Biol,2013,8(5):432~439.
11 Hoffman R M. Gene and stem cell therapy of the hair follicle[J]. Methods Mol Biol,2005,289:437~448.
12 Hsu Y C,Pasolli H A,Fuchs E. Dynamics between stem cells,niche,and progeny in the hair follicle [J]. Cell,2011(144):92~105.
13 Jaks V,Barker N,Kasper M,et al. Lgr5 marks cycling,yet long-lived,hair follicle stem cells[J].Nature Genetics,2008,40(11):1291~1299.
14 Jensen K B,Collins C A,Nascimento E,et al. Lrig1 expression defines a distinct multipotent stem cell population in mammalian epidermis[J].Cell Stem Cell,2009,4(5):472~439.
15 Jones P H,Watt F M. Separation of human epidermal stem cells for transit amplifying cells on the basis of differences in integrin function and expression[J]. Cell,1993,73:713~724.
16 Kloepper J E,Tiede S,Brinckmann J,et al. Immunophenotyping of the human bulge region:The quest to define useful in situ markers for human epithelial hair follicle stem cells and their niche[J]. Exp Dermatol,2008,17(7):592~609.
17 Lako M,Armstrong L,Cairns P M,et al. Hair follicle dermal cells repopulate the mouse heamatopoietic system[J]. J Cell Sci,2002,115:3967~3974.
18 Li L,Mignone J,Yang M,et al. Nestin expression in hair follicle sheath progenitor cells[J]. Proc Nati Acad Sci USA,2003,100(17):9958~9961.
19 Liu X,Song L,Liu J,et al. miR-18b inhibits TGF-β1-induced differentiation of hair follicle stem cells into smooth muscle cells by targeting SMAD2[J]. Biochem Biophys Res Commun,2013,438(3):551~556.
20 Owen T A,Aronow M,Shalhoub V,et al. Progressive development of the rat osteoblast phenotype in vitro:Reciprocal relationships in expression of genes associated with osteoblast proliferation and differentiation during formation of the bone extracellular matrix[J]. J Cell Physiol,1990,143:4204~4230.
21 Owens P,Han G,Li A G,et al. The role of Smads in skin development[J]. Invest Dermatol,2008,128(4):783~790.
22 Raucci F,Di Fiore M M. Localization of c-kit and stem cell factor (SCF) in ovarian follicular epithelium of a lizard,Podarcis s. sicula[J]. Acta Histochem,2011,113(6):647~655.
23 Snippert H J,Haegebarth A,Kasper M,et al. Lgr6 marks stem cells in the hair follicle that generate all cell lineages of the skin[J]. Science,2010,327(5971):1385~1389.
24 Sottile V,Thomson A,McWhir J. In vitro osteogenic differentiation of human ES cells[J]. Cloning Stem Cells,2003,5(2):149~155.
25 Terunuma M O. Characterization and isolation of stem cell-enriched human hair follicle bulge cells [J]. J Clin Invest,2006,116:249~260.
26 Toma J G,Akhavan M,Fernandes J L. Isolation of multipotent adult stem cells from the dermis of mammalian skin[J]. Nature Cell Biology,2001,3:778~784.
27 Xu Z C,Zhang Q,Li H. Differentiation of human hair follicle stem cells into endothelial cells induced by vascular endothelial and basic fibroblast growth factors[J]. Mol Med Rep,2014,9(1):204~210.
28 Yang J S,Lavker R M,Sun T T. Upper human hair follicle contains a subpopulation of keratinocytes with superior in vitro proliferative potential [J]. J Invest Dermatol,1993,101(5):652~659.

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Isolation and Cultures of Hair Follicle Stem Cell from Cashmere Goat and Identification of Induced Differentiation

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