Dispersed Cell Culture of Human Sweat Duct Cells under Serum‐free Conditions
Dispersed Cell Culture of Human Sweat Duct Cells under Serum‐free Conditions
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DOI：
10.1111/j.1346-8138.1993.tb01364.x
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发表时间：
1993-11
期刊：
影响因子：
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通讯作者：
N. Uchida;H. Oura;H. Nakanishi;Y. Urano;S. Arase
中科院分区：
文献类型：
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作者：
N. Uchida;H. Oura;H. Nakanishi;Y. Urano;S. Arase
使用无血清培养基K-GM培养基成功培养了人小汗腺导管细胞。从经分散酶处理的来自手掌或脚底的皮肤样本分离外分泌汗管。用胰蛋白酶和EDTA处理分离的导管后，在K-GM培养基中培养分散的细胞。在原代培养物中，接种后3至4天观察到小菌落。然后细胞迅速增殖并形成具有铺路石样细胞排列的大集落。在培养过程中，有时在菌落中心形成小的圆顶状区域。培养物最多繁殖7代。第1 ~ 6代细胞的植板率约为20%~ 30%。免疫细胞化学显示，培养的细胞与抗癌胚抗原K8.37和K8.13呈阳性染色，但与抗S100蛋白、抗HLA-DR、34βB4或PKK 3不呈阳性染色。培养细胞的电子显微镜照片显示由桥粒连接的多层扁平细胞。这些结果表明，培养的细胞具有与外泌汗腺导管部分的染色特性相容的染色特性。在培养过程中未观察到其他间充质细胞（如成纤维细胞）的污染。
Human eccrine gland duct cells were successfully cultured using a serum‐free medium, K‐GM medium. Eccrine sweat ducts were isolated from dispase treated skin specimens from palms or soles. After treatment of the isolated ducts with trypsin and EDTA, dispersed cells were cultured in K‐GM medium. In primary cultures, small colonies were seen 3 to 4 days after inoculation. Then the cells rapidly proliferated and formed large colonies with a paving stone‐like cell arrangement. During the culture, small dome shaped areas were sometimes formed in the centers of colonies. Cultures multiplied for a maximum of 7 passages. The plating efficiencies of the 1st to 6th passage cells were about 20% to 30%. Immunocytochemically, cultured cells were positively stained with anti‐carcinoembryonic antigens, K8.37 and K8.13, but not with anti‐S100 protein, anti‐HLA‐DR, 34βB4, or PKK3. An electron micrograph of the cultured cells showed a multilayer of flattened cells linked by desmosomes. These results indicate that the cultured cells possessed the staining properties compatible with those of the ductal portion of eccrine sweat glands. No contamination by other mesenchymal cells, such as fibroblasts, was seen during the culture.