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Exploration and Research on the Reconstruction of Hair Follicle by in Vivo and Vitro

Author: LiKunJie
Tutor: HeLi；LiuLing
School: Kunming Medical College
Course: Dermatology and Venereology
Keywords: Cell culture Outer root sheath cells Dermal papilla cells Tissue Engineering Sodium alginate bracket
CLC: R329
Type: Master's thesis
Year: 2011
Downloads: 23
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Abstract

Objective: Isolation and culture of hair follicle-derived seed cells in vitro and in vivo induction of hair follicle formation, and to explore the role of the biological characteristics of the hair follicle and seeds cells in the hair follicle formation. Methods: (1) in combination with enzyme digestion tissue culture obtained from neonatal foreskin dermal fibroblasts (Fb); (2) the purchaser of the hair follicle outer root sheath the cell lines (HHFORSC) and the human hair follicle dermal papilla cell lines ( HHDPC), parallel recovery, culture and subculture; (3) in the outer root sheath cells (ORSC), after the intervention of the dermal papilla cells (DPC) and mitomycin fibroblast cell suspension by a certain combination and sodium alginate 3D cell scaffold seeded in different vaccination order to build three-dimensional model of the hair follicles in vitro cultured for 8 weeks, HE staining observed by light microscopy of hair follicle formation; at the same time the three-dimensional model transplanted into the bal / bcl nude mice subcutaneous 8 weeks, feeding nude HE staining, immunohistochemistry and electron microscopy of hair follicles formed line drawn at the transplant site. Results: (1) successfully get people FB; recovery (2), cultured and passaged HHFORSC, HHDPC and self-separation, cultured Fb growth; (3) build in vitro hair follicle three-dimensional model, HE staining visible bracket have uniformly scattered in distribution of hair follicle cells, did not see the keratinized material and hair follicle structure; (4) nude mice graft HE staining cells clump together, the hair follicle-like structures were arranged in a circle, CK14, CK15, beta1 integration hormone and vimentin positive staining; visible in the electron microscope, model attached to the bracket on the hair follicle cells and red blood cells. (5) The first seeded with mitomycin intervention fibroblast 1 × 103 in cultured week after inoculation DPC: ORSC (1:5) cell suspension in sodium alginate 3D cell stent graft HE staining nude mice not only seen clearly in the hair follicle-like structures formed, and the formation of a large number of hair follicle-like structure; (6) MTT concentration of 10ng/ml bone morphogenetic protein 4 (BMP-4) dermal papilla cells and outer root sheath cells promote value-added; (7) seed cells in vitro medium KGM group hair follicle cells to aggregate into clumps of cells, some arranged in a ring, with the the MSCM group-like structure of the cell mass, no ring structure. Conclusion: the enzyme digestion and tissue culture method used in this test Fb maintained a good growth characteristics; hair follicles in vitro three-dimensional model of culture failed to induce hair follicle-like structure, but in the bal / bcl nude mice in vivo induced hair follicle samples structure, and the model DPC maintain the ability to induce hair follicle formation, ORSC maintain the characteristics of the hair follicle stem cells. In addition, the trial to explore the combination of the best cells induce hair follicle formation, the order of inoculation and the proportion of first inoculated with mitomycin intervention fibroblasts 1 × 103 sodium alginate 3D cell scaffold, training a week after the vaccination DPC: ORSC (1:5) cell suspension; also confirmed that the concentration of 10ng/ml BMP-4 can significantly promote the the DPC and ORSC the proliferation; seed cells in vitro significantly better than MSCM medium KGM. This test pointed out the direction for the successful reconstruction of the hair follicle.

Exploration and Research on the Reconstruction of Hair Follicle by in Vivo and Vitro

Abstract

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