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.     

人毛乳头细胞在不同传代时期某些细胞因子表达的变化

目的 观察毛乳头细胞在体外培养条件下的生长特性,为毛囊重建提供参考资料.方法 采用二步酶消化法分离正常人头皮毛囊的毛乳头细胞,在体外进行传代培养,用免疫组化方法观察不同传代毛乳头细胞生长因子表达的差异.结果 高代培养毛乳头细胞逐渐丧失其凝集性生长的特性,生长因子的表达逐渐消失.结论 毛乳头细胞的生长及其特性的保持受许多因素的影响,体外培养的毛乳头细胞与体内有差异.低代毛乳头条件培养基可恢复毛乳头细胞部分生长特性.     

毛乳头细胞凝集性生长对诱导毛囊样结构形成能力的影响

目的　以细胞外基质成分纤维连接蛋白促进培养的人头皮毛乳头细胞凝集性生长行为 ,观察毛乳头细胞的凝集性生长对诱导毛囊形成能力的影响。方法　传代培养的人头皮毛乳头细胞经细胞外基质纤维连接蛋白预处理后 ,促进形成凝集性生长团 ,再与毛囊上皮细胞共同移植于裸鼠皮下 ,分别于 8、12周进行组织学检查 ,观察毛囊形成及分化程度。结果　 8周后纤维连接蛋白预处理的毛乳头细胞组可见有毛囊样结构形成 ,未处理组仅见松散细胞团 ;12周后两组均可见有毛囊样结构形成 ,但纤维连接蛋白预处理组较未处理组诱导形成毛囊样结构分化更成熟 ;真皮成纤维细胞组未见有毛囊样结构形成。结论　毛乳头细胞的凝集性生长特性对其诱导毛囊形成的能力有密切的关系 ,细胞呈凝集性生长后 ,其生物学功能增强。     

毛乳头细胞微囊诱导大鼠耳毛囊再生

目的:应用人头皮毛乳头细胞APA(alginate-polylysine-alginate,海藻酸钠-多聚赖氨酸-海藻酸钠)微囊诱导大鼠耳部毛囊再生。方法:以APA微囊包裹体外分离培养的人头皮毛乳头细胞;将毛乳头细胞微囊移植至大鼠耳部皮下,定时行组织学检查。结果:毛乳头细胞微囊移植3周后,组织学检查显示:移植部位皮下形成有密集的同心圆状毛囊结构形成,其数量、形态、分化程度等与正常鼠耳毛囊明显不同。结论:微囊化毛乳头细胞具备诱导大鼠耳部毛囊再生的功能。     

毛囊细胞混合物移植诱导裸鼠毛囊样结构形成

目的 观察毛囊细胞复合物移植诱导裸鼠毛发再生和毛囊重建情况.方法 采用裸鼠移植技术,将混合分离出来的毛囊细胞,包括毛囊毛乳头细胞、毛囊外根鞘细胞、毛囊真皮鞘细胞和毛囊真皮成纤维细胞等,移植到裸鼠皮下,观察毛囊形成情况.结果 在裸鼠的皮肤切片中可以看到较为完整的毛囊结构形成.结论 毛囊细胞混合物可以在体内诱导出毛囊样结构形成.     

成年鼠被毛毛乳头细胞分离方法的建立与评价

目的 寻求一种简单高效的分离培养小鼠被毛毛乳头细胞的方法.方法 取5周龄C57小鼠背部皮肤,刮去毛发后予0.2％Ⅰ型胶原酶37℃消化2h,刮下毛球部,经过2次Ficoll密度梯度离心后,得到纯净的毛乳头进行培养.检测毛乳头贴壁率、毛乳头细胞特异性标记表达及其体内毛囊重建能力.结果 该分离方法能显著降低工作强度,减少污染机会,获得的毛乳头贴壁快、细胞迁出快.qRT-PCR、细胞免疫荧光实验结果均证实体外培养的小鼠被毛毛乳头细胞可表达与毛囊诱导能力相关的特异性标记物ALP、β-catenin及Versican,且与触须毛乳头细胞相比差异无统计学意义(P＞0.05).体外培养的小鼠被毛毛乳头细胞具有诱导毛囊再生的能力.结论 胶原酶消化结合Ficoll梯度离心是一种简单、有效地分离获取小鼠背部毛乳头的方法.     

影响毛囊器官型培养中胶原凝胶收缩的几个因素分析

目的 :观察鼠尾胶原、毛乳头细胞和毛囊上皮细胞的浓度对毛囊器官型培养模型中胶原凝胶收缩的影响。方法 :用不同浓度的鼠尾胶原、毛乳头细胞和毛囊上皮细胞制备成毛囊器官型培养模型 ,体外培养 10 d,隔日测量培养凝胶的直径 ,观察其对胶原凝胶收缩的影响。结果 :鼠尾胶原、毛乳头细胞和毛囊上皮细胞的浓度对毛囊器官型培养模型中胶原凝胶的收缩均有明显影响 (P<0 .0 1)。鼠尾胶原的浓度越高 ,胶原凝胶的收缩程度越小 ;而毛乳头细胞和毛囊上皮细胞的浓度对胶原凝胶收缩的影响则恰好相反 ,浓度越高 ,胶原凝胶的收缩程度越大。结论 :鼠尾胶原、毛乳头细胞和毛囊上皮细胞的浓度是影响毛囊器官型培养模型中胶原凝胶收缩的主要因素。     

人头皮毛乳头细胞体内诱导毛囊形成的实验研究

目的 探讨人头皮毛乳头细胞和毛囊上皮细胞在体内的相互作用，了解毛乳头细胞在体内诱导毛囊形成和调控毛囊生长发育的能力，为毛囊细胞移植奠定实验基础。方法 人头皮毛乳头细胞和毛囊上皮细胞共同移植到无胸腺裸鼠体内，在不同时相取材进行组织切片。H—E、角蛋白免疫组织化学染色。结果 毛乳头细胞和毛囊上皮细胞在体内相互作用，首先形成不规则的混合细胞团、逐渐发展到排列规则、最终形成完整的毛囊结构，并且在移植腔上方的裸鼠皮肤内形成了毛囊结构。这些结构表达毛囊特有的角蛋白。结论 培养的毛乳头细胞与毛囊上皮细胞在裸鼠体内能形成完整的毛囊结构，同时也能诱导裸鼠角质形成细胞、形成毛囊结构。     

双氢睾酮体外培养人毛囊中差异表达的miR-133b对人毛乳头细胞的增殖及诱导能力的影响

探究不同浓度双氢睾酮（DHT）对毛囊生长和毛囊细胞增殖的影响及其与miR-133b表达的关系,然后进一步探究miR-133b对毛乳头细胞的增殖和诱导能力的影响。     

诱导多能干细胞治疗脱发研究新进展

脱发性疾病是临床上常见的一组影响美容、心理和健康的疾病。目前,脱发性疾病的发病机制仍不甚清楚。毛囊结构、毛乳头细胞及毛囊干细胞异常可能与脱发性疾病密切相关。诱导多能干细胞(i PSC)是将外源性转录因子导入已分化的体细胞中并重新编排形成的具有类似于胚胎干细胞全能性的多潜能细胞,在临床医学和再生医学方面具有广阔的应用前景。i PSC可分化为毛乳头细胞、毛囊干细胞及再生毛囊,为治疗脱发性疾病开辟了新道路。     

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.     

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.     

毛囊球部细胞体外在胶原/壳聚糖多孔支架上形成皮肤样结构的研究

目的:观察毛囊球部细胞种植到胶原/壳聚糖多孔支架上皮肤重建情况.方法:采用细胞培养技术和细胞支架种植技术,将培养的毛囊球部细胞种植到胶原/壳聚糖多孔支架上,HE染色观察皮肤样结构形成情况,并作免疫组织化学鉴定,以证明细胞的来源属性.结果:毛囊球部细胞种植到胶原/壳聚糖多孔支架上可见皮肤样结构形成,表皮部分细胞分化良好,排列复层整齐,有角化现象,高分子与低分子角蛋白表达阳性.结论:低传代培养的毛囊球部细胞在体外胶原/壳聚糖多孔支架上可诱导皮肤样结构形成.     

Expression of stem cell factor in hair follicl epithelium

Thedermalepidernlalinteractioninthehairfollicleisahotspotincytobiologlcalstudies.anytoklnesplayimportantrolesinin(luctionofbiologicafactivitiesbetweentilegermalandepidermal].Stemcellfactor(St'F).whichwasfoundinrecentyears.isacyloklnerelatedicthestir\-tval.growl11anddevelopmentofthehemop()leticstemcellsandcanexertimportanlbiologicaleffectsonthedevelopmentofkeratlnocytesandnlelanocytes"/'.Inthisstudy.theexpressionofSCFInhumanhairfollicleepitheliumwasin\'estigateclwithimmunohistochemistryandI)…     

ET-1、rhEGF、bFGF对培养的兔角膜内皮细胞的影响

目的 研究内皮素-1(ET-1)、重组人表皮生长因子(rhEGF)、碱性成纤维细胞生长因子(bFGF)对培养的兔角膜内皮细胞增殖能力的影响及其相互作用。方法 在体外培养的兔角膜内皮细胞中单独使用或联合应用ET-1、rhEGF、bFGF，采用MTT方法观察对细胞增殖的影响，免疫组化染色法和计算机图像分析系统检测对细胞增殖细胞核抗原(PcNA)表达的影响。结果 一定浓度ET-1促进培养的兔角膜内皮细胞的增殖，且呈剂量相关性。10pmol／L时起作用，200pmol／L时发挥最大作用。10ng／ml rhEGF、2ng／mlbFGF也促进培养的兔角膜内皮细胞的增殖。联合应用ET-1 rhEGF、ET-1 bFGF、ET-1 rhEGF bFGF，细胞吸光度(A)值明显高于单独使用相应药物时A值之和。ET-1、rhEGF、bFGF单独使用可促进培养的兔角膜内皮细胞PCNA表达，联合应用时PCNA表达平均吸光度A值明显强于单独使用相应药物时表达强度之和。结论 ET-1可作为一种生长因子，它和rhEGF、bFGF单独使用可促进培养的兔角膜内皮细胞增殖能力，联合应用时具有协同作用。     

毛乳头细胞的研究现状

毛囊是一种皮肤附属器官,其结构的完整不仅为机体新陈代谢所需,同时对于动物的取暖、人的外表美观都有重要意义。毛囊具有自我更新和周期性生长的特点,这与毛球部存在的一群间充质细胞密切相关,即毛乳头细胞(DPC),它在毛囊发育以及生后毛囊周期过程中都发挥着重要的诱导作用。缺失DPC,无法形成毛囊;而在生后毛囊周期中,DPC通过与周围的毛囊上皮细胞相互作用,直接或间接地诱导毛囊周期性的生长。研究DPC的特性及其发挥的生物学功能,对了解毛囊的发育和再生具有重要的指导意义。     

毛乳头细胞的培养与鉴定

目的：建立毛乳头细胞的培养方法,以进一步研究毛乳头细胞在毛发再生中的作用和意义．方法：分离得到完整毛囊,依次采用Dispase酶和胶原酶消化得到毛乳头,再进行毛乳头细胞的培养．结果：成功进行了毛乳头细胞的原代和传代培养,所获细胞经细胞化学染色证实细胞具有明显的异染特性．毛乳头贴壁后5～6d周围即可见细胞长出,细胞生长快,12～16d左右细胞即可融合．在倒置显微镜下细胞呈成纤维细胞样,有聚集生长特性,至少可以传15代．冻存后复苏细胞贴壁率在80％以上．结论：该方法是体外培养毛乳头细胞的理想方法,操作较为简单,降低了工作强度,所获细胞纯度高,可以长期传代、冻存．