酶消化法联合组织块法与经典酶消化法体外分离培养黑色素细胞的比较  

Melanocytes isolation in vitro by enzyme digestion combined with tissue block or enzyme digestion

作　　者：牛牧[1,2] 李其林[1,2] 何丹华[1,2] 黄永华[1,2] 陆小娟[1,2] 戴丽冰 

机构地区：[1]暨南大学第四附属医院 [2]广东省广州市红十字会医院皮肤科,510220 [3]广东省广州市创伤外科研究所,510220

出　　处：《广东医学》2013年第4期501-504,共4页Guangdong Medical Journal

基　　金：广东省自然科学基金资助项目(编号:9151051501000006)

摘　　要：目的对酶消化法联合组织块法(以下简称"联合法")与单纯酶消化法体外分离培养人黑色素细胞进行比较,以找到更加快速、经济、简便的黑色素细胞体外分离培养方法。方法使用倒置相差显微镜连续多代观察两种方法体外培养的黑色素细胞原代、传代情况,分别从形态学、生长情况等方面对比两种方法培养黑色素细胞的差别。结果细胞形态:联合法分离的细胞同源性好,细胞较细长,树突较多且相互交叉成网状呈集落分布,但初代培养时间较长;酶消化法分离的细胞同源性一般,初代细胞较粗短,树突较少且细胞分布较分散,但初代培养时间短。两种方法细胞形态随着传代次数的增多渐趋一致。生长情况:酶消化法分离的细胞一次可获得较大的细胞量,但容易污染,且初代培养的细胞混有较多的上皮细胞,需多次传代纯化;联合法分离的细胞较单一,但细胞量相对较少,两种方法分离细胞增殖和生长曲线大体一致。结论联合法可获得比较可观的细胞量,两次传代后即可获得比较纯净的黑色素细胞,可避免胰蛋白酶对黑色素细胞的损伤,是较为快速、经济的黑色素细胞体外分离方法。Objective Human melanocytes were isolated and cultured by enzyme digestion combined with tissue block （hereinafter referred to as ＂the combine method ＂ ） and simple enzyme digestion in vitro. Two methods were cornpared, in order to investigate a more rapid, economic, and simple culture method of human melanocytes. Methods Melanocytes cultured by two methods were continuously observed by using the inverted microscope, for comparison of the cellmorphology, proliferation, and so on. Results The melanocytes isolated by the combined method presented with fine homogeneity, which were slender, with more dendrites and intersected the mesh in colony distribution, though it took Ion-ger for primary culture. That was in contrast with enzymatic digestion method. A morphological consistency was observed with the 2 methods as generating. Although more cells were obtained isolated by enzymatic digestion method, they wereeasily contaminated, and required further multiple purification due to epithelial cell mixture. That was in contrast with the combined method. The proliferation and growth curves obtained by these two methods were consistent. Conclusion Thecombine method can obtain relatively substantial and pure melanocytes after two passages protect cells from trypsin digestion, providing a rapid, economic, and simple culturing method for human melanocytes in vitro.

关 键 词：黑色素细胞 联合法 酶消化法 体外培养 

分 类 号：R783.5[医药卫生—口腔医学]