胎儿头皮毛囊内黑素细胞HMB-45和NKI/beteb的表达(1)
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[摘要]目的:观察黑素细胞在胎儿头皮内的定位。方法:获取胎儿头皮(胎龄7月,畸形引产,死亡2h),制备冰冻切片,分别用HMB-45和NKI/beteb抗体进行免疫荧光染色和免疫组化染色,激光共聚焦显微镜和倒置显微镜观察摄片。结果:NKI/beteb阳性细胞主要位于胎儿头皮的毛囊外根鞘隆突区,也见于毛球处、表皮基底部。HMB-45阳性细胞主要存在于毛球部、毛母质和表皮基底部,外根鞘区表达则阴性。结论:胎儿头皮毛囊内存在不同状态的黑素细胞,位于毛囊外根鞘隆突区的黑素细胞是黑素母细胞。
[关键词]毛囊;胎儿;隆突;黑素细胞
[中图分类号]Q813.1 [文献标识码]A [文章编号]1008-6455(2010)07-1003-04
Location of melanocytes in hair follicle of fetal scalp
LI Hong-zhou,XUE Dong-zhang,ZHANG Ru-zhi,ZHU Wen-yuan,ZHANG Jing,CHEN Ze-nong,ZHAO Hao
(1. Department of Dermatology,First Affiliated Hospital of Bengbu Medical University, Hefei 233004,Anhui,China)
Abstract:ObjectiveTo explore the location of melanocytes in hair follicle of fetal scalp. MethodsThe scalp was from a died fetus (fetal age of 7 months, induction delivery due to developmental abnormity ) and prepared for frozen sections, thenwhich respectively received immunofluorescent and immunofluochemical staining with HMB-45 and NKI/beteb antibodies. The stain results was observed and photographed by confocal microscope and the inverted microscope, respectively. ResultsThe positive cells stained by NKI/beteb antibody were mainly located at the bulge regions in the outer root sheath (ORS) of hair follicle, some in the hair bulbs and the epidermal basilar parts. The HMB-45 positive cells were present in the hair bulb, the hair matrix and the epidermal basilar parts, but not in ORS.ConclusionThere are melanocytes with different status in hair follicle. Melanocyte at the bulges of ORS is melanoblast.
Key words:hair follicle; fetus; bulge; melabocyte
毛囊是一个独特的皮肤附属器, 其色素单位的自我更新取决于毛囊内的黑素干细胞(melanocyte stem cells, MSCs)及其子代细胞。毛囊MSCs特征如下:具备不成熟、慢循环、自我修复、产生子代细胞,并且活化增殖毛发生长早期的细胞。毛囊黑素干细胞 (follicle melanocyte stem cells,FMSC)是MSCs中的一种。有报道鼠毛囊外隆突区检测出MSCs[1]。鼠的胚胎研究显示,FMSC存在于毛囊永久性区域的下部毛囊隆突区,具有MSCs的特征,受刺激后可以分化成为成熟的黑色素细胞[2]。因此,笔者设计此实验初步定位人胎儿的FMC,观察HMB45,NKI/beteb在人类胎儿毛囊中的表达情况。
1材料和方法
1.1 材料
1.1.1 标本来源:引产胎儿的头皮(胎龄7月,畸形引产),取标本时,胎儿已死亡2h。
1.1.2主要仪器及试剂:小鼠抗人HMB-45单克隆抗体(北京中山生物技术公司);小鼠抗人NKI/beteb单克隆抗体(Abcam);DyLightTM488标记山羊抗小鼠IgG(北京中山生物技术公司)辣根酶标记山羊抗小鼠IgG(北京中山生物技术公司);OLYPUS倒置显微镜;激光共聚焦显微镜(NIKON-EZ-C1SI)。
1.2 方法
1.2.1 标本处理:获取胎儿头皮,用碘伏浸洗5min后,酒精去碘伏,接着生理盐水冲洗数次,剪刀修剪组织标本,去除脂肪,筋膜等多余组织,再用生理盐水冲洗数次,最后多聚甲醛固定组织15min后,制备冰冻切片(4μm),用冷丙酮固定15min ......
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