人游离毛囊体外培养及动态观察

目的 进行人发毛囊的体外培养，并对其中后期改变进行形态学、组织学观察。方法将美容除皱术中切除的头皮在无菌条件下分离出游离的毛囊，培养于含血清DMEM培养基中，动态观测毛囊的生长速度及形态学、组织学改变。结果 成功进行了人发毛囊的体外培养，毛囊培养中后期呈退化期改变。结论 在含血清DMEM培养基中生长的毛囊易较早进入退化期，简化了游离毛囊的常规病理制片方法。     

章光101生发系列产品对C57BL/6J小鼠触须毛囊体外培养的研究

目的 应用C57BL/6J小鼠触须毛囊体外培养模型,观察章光101生发系列产品对毛囊生长的影响.方法 分离小鼠触须毛囊,放人Williams'E无血清培养基中,将毛囊分为4组,分别为空白对照组,毛囊滋养液组、防脱育发剂组和三参头发宝组,观察毛囊的形态变化、生长长度并检测凋亡细胞.结果 毛囊培养第8 d,对照组毛囊毛发生长长度为(0.669±0.32)mm,毛囊滋养液组为(1.312±0.47)mm,防脱育发剂组为(1.345±0.51)mm,三参头发宝组为(1.285±0.38)mm,3个实验组均快于对照组(P<0.05),但3个实验组之间差异不显著;毛球形态观察,对照组毛囊毛球体积变小,结构疏松,出现退行性变化,而实验组毛囊毛球形态较圆,结构致密,仍处于生长期;TUNEL法检测毛囊中的凋亡细胞,3个实验组和对照组平均每个毛囊中的凋亡细胞数分别为6.58±1.08,5.83±1.12,6.92±1.24和21.92±2.11,对照组中的凋亡细胞数明显高于实验组(P<0.05),但3个实验组之间差异不显著.结论 章光101生发系列产品能促进小鼠触须毛囊的生长,延缓其进入退行期.     

雄激素受体在大鼠触须毛囊干细胞中的表达研究

目的探讨在毛囊周期的3个不同时期(生长期、退化期和静止期)中大鼠毛囊干细胞中以及体外培养的毛囊干细胞中雄激素受体(androgen receptor,AR)的表达规律及其意义.方法采用免疫组织化学方法检测AR表达,采用总RNA提取法及RT-PCR检测AR mRNA的表达情况.结果①在体情况下,生长期、退化期和静止期毛囊干细胞中均有AR的表达,AR mRNA在生长期呈强表达,而在退化期和静止期表达微弱;②体外情况下,培养的毛囊干细胞中有AR的表达.结论毛囊干细胞在整个毛囊周期中均有AR的表达,是雄激素作用的靶细胞;AR mRNA在毛囊周期中呈规律性表达,提示雄激素与其受体的相互作用可能对毛囊干细胞的分化具有重要的促进作用.     

Pax6基因和蛋白在大鼠触须毛囊中的表达

目的 初步研究转录因子Pax6(paired homebox 6)在大鼠触须毛囊中的表达.方法 运用免疫组织化学法检测Pax6蛋白在大鼠触须毛囊及培养的毛囊bulge细胞巾的表达,RT-PCR分析毛囊中Pax6两种亚型的表达情况.结果 大鼠触须毛囊细胞巾Pax6表达阳性,RT-PCR显示表达量以Pax6-5a为高,在体外培养细胞中表现出细胞增殖能力与Pax6在细胞中的表达部位相关.结论 Pax6在大鼠触须毛囊中表达,可能在其形成和功能的维持上发挥一定的作用.     

横断毛囊体外再生的形态学研究

目的：研究毛囊横断后再生的条件，探索毛囊干细胞的初步定位．方法：将人头皮分离成游离的毛囊，共进行了8批横断毛囊培养，每批培养毛囊约30根，每批分3组，从不同位置(紧贴毛球、毛囊下1／3处、毛囊下1／2处)横断毛囊后进行培养，观测毛囊的生长情况及形态学改变．结果：紧贴毛球上方或在毛囊下1／3横断，上段毛囊大部分可以再生出新毛球样结构，毛囊可继续生长，上段毛囊平均生长长度分别为0．78mm和0．67mm，可生长比例分别为75％和70％；在毛囊下1／2横断，上段毛囊可生长比例仅为12．5％，明显低于前两组(P＜0．01)．结论：在一定位置横断毛囊，上段的毛囊细胞可以发生增殖分化，毛囊可以继续生长，并再生出新的毛球样结构，表明上段毛囊是其干细胞所在地．     

大鼠生长期触须毛囊干细胞中FGF-13的表达研究

目的探讨大鼠生长期触须毛囊中以及体外培养的毛囊隆突部细胞即毛囊干细胞中FGF-13的表达分布及其意义.方法分别采用免疫组织化学方法和免疫细胞化学方法检测FGF-13的表达.结果体外和在体两种情况下,毛囊隆突部细胞都有FGF-13的表达,在体情况下毛囊Bulge区以下毛球部以上的外根鞘细胞中亦有干细胞标记和FGF-13的表达.结论提示FGF-13参与生长期触须毛囊隆突部细胞沿毛囊外根鞘向下迁移这一过程.     

人游离毛囊体外培养及动态观察

目的进行人发毛囊的体外培养,并对其中后期改变进行形态学、组织学观察。方法将美容除皱术中切除的头皮在无菌条件下分离出游离的毛囊,培养于含血清DMEM培养基中,动态观测毛囊的生长速度及形态学、组织学改变。结果成功进行了人发毛囊的体外培养,毛囊培养中后期呈退化期改变。结论在含血清DMEM培养基中生长的毛囊易较早进入退化期,简化了游离毛囊的常规病理制片方法。     

螺内酯对体外培养的人毛囊影响的实验研究

目的：观察螺内酯（spironllactone）对体外培养的人毛囊生长的影响。方法：建立人毛囊体外培养模型，加入不同浓度的螺内酯，通过目镜测微尺测量毛囊的长度，利用^3H-胸腺嘧啶核苷酸（^3H-TdR）掺入法检测毛囊DNA合成率。结果：螺内酯各浓度组毛囊生长长度与空白对照组相比无明显差异（P〉0．05）。^3H-TdR掺入法检测与长度检测结果一致。结论：螺内酯对体外培养的人毛囊的生长无促进和抑制作用。     

毛囊的体外保存和延期移植

在各种以毛发移植治疗男性型秃发的手术中，使用毛囊单位作为移植物是目前较理想的方法，但它在手术人员数量和设备上提出了更高的要求。延期毛发移植将允许把一次任务繁重的移植手术拆分为两个独立的步骤，在特定情况下有一定的实用意义。本文着重对毛囊单位体外保存和延期移植研究的意义、影响体外毛囊活性的主要因素、以及毛囊活性常用评估方法等方面作一综述。     

自制毛囊再生乳膏对小鼠皮肤创面毛囊再生的影响

背景 创面愈合时无毛囊等皮肤附属器再生是创面修复领域的难题.小分子化合物能够调控细胞增殖与分化,促使成熟体细胞再分化为具有增殖能力的前体细胞或其他细胞系,还能够靶向调控信号通路与代谢进程,是一种促进皮肤及附属器原位再生的安全有效的方法.目的 本研究拟通过小分子化合物组合探索其促进创面毛囊再生的效果及机制.方法 选取C5...     

胎鼠表皮干细胞的分离培养及毛囊再生

0　引言　近年来 ,随着组织工程的飞速发展 ,人类组织干细胞的研究及其临床应用日益成为国内外学者关注的焦点 ,表皮干细胞即是其中之一 .因此 ,表皮干细胞的培养成功为利用组织工程原理及分子生物学手段构建全新的人工皮肤组织带来了希望 .1　材料和方法1.1　材料　ＤＭＥＭ培养基、胰蛋白酶、ｄｉｓｐａｓｅ酶及角朊细胞培养基均购自Ｇｉｂｃｏ公司 ,小鼠ＩＶ型胶原购自Ｓｉｇｍａ公司 ,ｉｎｔｅｒｇｒｉｎ β1兔多抗为ＳａｎｔａＣｒｕｔｚ公司产品 ,一级昆明小鼠由第四军医大学实验动物中心提供 ,裸鼠购自上海生物所 .1.2　方法1.2 .1　…     

毛囊细胞移植诱导裸鼠毛囊样结构形成的研究

目的:观察毛囊细胞移植诱导裸鼠毛发再生和毛囊重建情况.方法:采用细胞培养技术和裸鼠移植技术,将培养的毛囊毛乳头细胞、真皮鞘细胞和头皮真皮成纤维细胞按比例与毛囊上皮细胞混合,移植到裸鼠皮下,观察毛囊形成情况.结果:在毛囊毛乳头细胞与毛囊上皮细胞混合后移植到裸鼠皮下后可见毛囊样结构形成,而毛囊真皮鞘细胞和头皮成纤维细胞与毛囊上皮细胞混合则不能诱导裸鼠毛囊样结构形成.结论:低传代培养的毛乳头细胞与毛囊上皮细胞混合后在体内可诱导毛囊样结构形成.     

Biological characterization of cultured dermal papilla cells and hair follicle regeneration in vitro and in vivo

Background Dermal papilla cells (DPC) are a group of mesenchyme-derived cells at the base of the hair follicle, where they regulate and control hair follicle growth through the expression and secretion of cytokines. Nevertheless, the role of DPC derived chemokines and other cytokines in the hair follicle biology remain speculative. In this study, we investigated the expression of basic fibroblast growth factor (bFGF), endothelin-1 (ET-1) and stem cell factor (SCF) in different passages of cultured DPC and their effects on the biological behaviour of DPC.Methods The expression of bFGF, ET-1 and SCF in different passages of cultured DPC and their possible effects on the biological behavior of DPC are investigated using in situ hybridization and immunochemistry. In addition, we performed transplantation of hair follicle cells into nude mice. The cultured DPC, dermal sheath cells and fibroblast of human scalp, respectively, were mixed with cells of the hair follicle epithelium in different ratios, and then were cultured in hair follicle organotypic cultures or implanted into the subcutis of nude mice.Results The expression of ET-1 and SCF in early passages of cultured DPC became stronger, but turned weaker and even negative in late passages (>6 passages). Hair follicle-like structures were formed after DPC combined with the cells of hair follicle epithelium cells in hair follicle organotypic cultures. When hair follicle organotypic cultures were implanted into the subcutis of nude mice, the relative intact hair follicles were formed. After the transplantation of hair follicle cells into the nude mice, the hair follicle-like structure was formed in the group that contained DPC mixed with hair follicle epithelium cells. However, no hair follicles were formed in the other two groups. It was found that the higher the expression of ET-1 and SCF in DPC, the stronger the ability of DPC to induce hair follicle regeneration.Conclusions The cultured DPC can induce hair follicle regeneration and sustain hair growth in vivo and in vitro. Moreover, the expression of ET-1 and SCF is correlated with the ability of DPC inducing hair follicle regeneration.     

Biological characterization of cultured dermal papilla cells and hair follicle regeneration in vitro and in vivo

Background Dermal papilla cells （DPC） are a group of mesenchyme-derived cells at the base of the hair follicle, where they regulate and control hair follicle growth through the expression and secretion of cytokines. Nevertheless, the role of DPC derived chemokines and other cytokines in the hair follicle biology remain speculative. In this study, we investigated the expression of basic fibroblast growth factor （bFGF）, endothelin-1 （ET-1） and stem cell factor （SCF） in different passages of cultured DPC and their effects on the biological behaviour of DPC. Methods The expression of bFGF, ET-1 and SCF in different passages of cultured DPC and their possible effects on the biological behavior of DPC are investigated using in sire hybridization and immunochemistry. In addition, we performed transplantation of hair follicle cells into nude mice. The cultured DPC, dermal sheath cells and fibroblast of human scalp, respectively, were mixed with cells of the hair follicle epithelium in different ratios, and then were cultured in hair follicle organotypic cultures or implanted into the subcutis of nude mice. Results The expression of ET-1 and SCF in early passages of cultured DPC became stronger, but turned weaker and even negative in late passages （〉6 passages）. Hair follicle-like structures were formed after DPC combined with the cells of hair follicle epithelium cells in hair follicle organotypic cultures. When hair follicle organotypic cultures were implanted into the subcutis of nude mice, the relative intact hair follicles were formed. After the transplantation of hair follicle cells into the nude mice, the hair follicle-like structure was formed in the group that contained DPC mixed with hair follicle epithelium cells. However, no hair follicles were formed in the other two groups. It was found that the higher the expression of ET-1 and SCF in DPC, the stronger the ability of DPC to induce hair follicle regeneration. Conclusions The cultured DPC can induce hair follicle regeneration and sustain hair growth in vivo and in vitro. Moreover, the expression of ET-1 and SCF is correlated with the ability of DPC inducing hair follicle regeneration.     

无血清K-SFM培养条件下大鼠毛囊Bulge细胞生物学特性的研究

目的研究无血清K-SFM培养条件下,大鼠毛囊Bulge细胞的生物学特性.方法分离大鼠毛囊Bulge细胞,分别置于无血清的K-SFM培养基(实验组)及有血清DMEM/F12培养基(对照组)的培养条件下培养.比较Bulge细胞在两种培养体系中的生长增殖和K19的表达状况.结果无血清培养的Bulge细胞在克隆形成率及K19的表达率均明显高于对照组(P＜0.05).结论体外培养和扩增大鼠毛囊Bulge细胞,无血清的K-SFM培养基较有血清DMEM/F12培养基更有利于Bulge细胞的扩增和表型的维持.     

大鼠皮肤及毛囊骨桥蛋白研究

目的 对出生和出生后大鼠皮肤及毛囊的骨桥蛋白（ｏｓｔｅｏｐｏｎｔｉｎ）的ｍＲＮＡ进行研究。方法 确定妊齿１４～１８ｄ和出生后１～６１ｄ Ｗｉｓｔａｒ大鼠毛周期,用放性同位素标记的原位杂交法牟大鼠皮肤和毛囊骨桥蛋白ｍＲＮＡ的分布和含量进行研究。结果 骨桥蛋白ｍＲＮＡ在大鼠出生后第１８天首次在真皮乳头（ｄｅｒｍａｌｐａｐｉｌｌａ,ＤＰ）细胞中出现,在各毛发周期中骨桥蛋白ｍＲＮＡ信号均出现一毛囊退化期Ｄ