摘要
目的研究geneticin在毛囊黑素细胞培养过程中对成纤维细胞的去除作用,研究12-O-十四烷佛波酯-13-醋酸酯TPA和含牛垂体提取物BPE角质形成细胞无血清培养基K-SFM对毛囊无色素黑素细胞AMMC形态和增殖的影响。方法通过胶原酶法培养毛囊黑素细胞,观察不同浓度geneticin对污染的成纤维细胞的去除。同时观察不同浓度TPA和含BPE的K-SFM对AMMC形态和促增殖的影响。结果采用100μg/mLgeneticin处理2d后,剩余细胞主要是黑素细胞,其中部分成纤维细胞呈死亡状态,继续培养至第7天后,黑素细胞的纯度达到90%以上。50ng/mLTPA可以促进细胞增殖,与100ng/mLTPA比较差异无显著性(P>0.05),但对细胞形态影响不大;100ng/mLTPA明显促进黑素细胞的树突增加。含20%、40%和80%的K-SFM含BPE培养基中,浓度为80%时促增殖作用最明显。结论100μg/mLgeneticin作用2d去除黑素细胞培养中污染的成纤维细胞是一种简单有效的方法,并可重复使用。在毛囊黑素细胞培养中,TPA以50ng/mL的浓度即可明显促进增殖,而不影响细胞的形态。含BPE的K-SFM可以浓度依赖性地促进AMMC增殖。
Objective:To investigate elimination of fibroblasts in cultures of hair follicle melanocytes from human scalp, and find out the effect of components in culture medium on the amelanotic melanocytes (AMMC). Methods:The method using collagenase V was employed to culture cells. Geneticin of different concentration was used to eliminate the contaminated fibroblasts. The transformation of cells was observed and the proliferation was tested by using different concentration TPA and K-SFM including BPE. Results:Treatment with geneticin of 100 μg/mL for two days eliminated the majority of fibroblasts. Rate of AMMC was more than 90 per cent 7 days after the treatment. 50 ng/mL TPA stimulated proliferation of the cells continuously and had no difference from 100 ng/mL TPA (P > 0.05). 100 ng/mL TPA induced rapid dentrite formation. Proliferation resulting from K-SFM including BPE depended on the concentration of BPE. Conclusions:Treatmenr with 100 μg/mL geneticin for two days can remove contaminated fibroblasts conveniently, and it can be used repeatedly. 50 ng/mL TPA is an appropriate concentration and it can stimulate proliferation of cells but can't cause transformation of AMMC. K-SFM including BPE stimulates the proliferation of AMMC in concentration-dependent manner.
出处
《临床皮肤科杂志》
CAS
CSCD
北大核心
2004年第2期80-83,共4页
Journal of Clinical Dermatology
基金
国家自然科学基金资助项目(30170861)
