高钙培养对表皮角质细胞形态和生长特点及角蛋白表达的影响  

Effects of high concentration of calcium in culture medium on morphology,growth characteristics and expression of keratin in human epidermal keratinocyte

作　　者：张翠萍[1] 付小兵[1] 李淑云[1] 孙同柱[1] 

机构地区：[1]解放军总医院第一附属医院全军创伤修复重点实验室,北京100037

出　　处：《感染．炎症．修复》2008年第2期87-89,F0002,共4页Infection Inflammation Repair

基　　金：国家重点基础研究发展规划项目(2005CB522603)

摘　　要：目的：观察高钙培养后表皮角质细胞的形态、生长特点及角蛋白表达的变化，进一步探讨钙在促进表皮角质细胞分化过程中的重要作用。方法：表皮角质细胞分别在低钙（0.05mmol/L）和高钙（1.2mmol/L）环境条件下培养1-2d，光镜下观察细胞的形态和生长特点，并采用免疫组织化学方法检测表皮角质细胞中角蛋白10（K10）、K14和K19的表达水平。结果：低钙培养的表皮角质细胞散在生长，分布均匀，细胞间隙较大，胞膜圆滑，边界清楚。高钙培养后，细胞胞体变得扁平，细胞间隙消失，并且细胞出现明显的成簇生长趋势。在正常培养即低钙培养条件下，K10和K19阳性细胞比例较少，所培养的细胞几乎全为K14阳性细胞；在高钙培养条件下，K10和K19阳性细胞比例明显增加，而K14的表达没有明显变化。结论：高钙培养能改变表皮角质细胞的形态和生长特点并提高细胞内角蛋白的表达水平，从而促进表皮角质细胞的分化成熟。Objective： To observe the changes in morphology, growth characteristic and expression of keratin in human epidermal keratinocytes cultured in medium with high-concentration of calcium, and to explore the important role of calcium in inducing the differentiation of epidermal keratinocytes. Methods： Human epidermal keratinocytes were cultured in low or high concentration of calcium （0.05 mmol/L or 1.2 mmol/L） for 1 to 2 days. The morphology and growth characteristic of epidermal keratinocytes were observed under microscope and the expression of keratin 10 （K10）, K14 and K19 was investigated by immunohistochemistry. Results： The human epidermal keratinocyte grew dispersively with equal distribution and large spaces between cells in culture with low calcium. The cell membrane was smooth with distinct border. After being cultured with high-concentration calcium, the morphology and growth characteristic of epidermal keratinocytes were obviously different. The cells became flattened with loss of intercellular space. In addition, there was aggregation of cells to from colonies. Immunohistochemistry showed that the expressions of K10 and K19 in epidermal keratinocytes cultured with high-concentration calcium were greatly enhanced, though there was no change in K14 compared with that in low-concentration calcium. Conclusion： Elevation of the calcium concentration in human keratinocyte culture increases the expression of keratins and induces the differentiation of epidermal keratinocytes.

关 键 词：表皮角质细胞 钙 分化 

分 类 号：R392-33[医药卫生—免疫学] R161[医药卫生—基础医学]