钙离子对体外培养皮肤角质形成细胞增殖与分化的调控作用

         作者：章培标，侯宜，侯立中，曲福军，周余来，颜炜群，杨同书

【摘要】  目的  观察无血清培养中不同钙离子浓度对皮肤角质形成细胞(Kc)增殖与分化的影响，为角质形成细胞的体外扩增和人工表皮形成提供依据。方法  取第二代新生兔背皮肤角质形成细胞，按0.5×104/cm2细胞密度接种于含不同浓度CaCl2角质形成细胞培养液的96孔板中，培养1周，每日观察细胞生长情况，至第7日以MTT法检测OD值。结果  当CaCl2浓度低于5mg/L时，细胞贴壁增殖缓慢，甚至无法贴壁；浓度为12.5mg/L 时Kc呈单层快速生长，并能保持较好的表型，是体外Kc培养扩增的最适钙离子浓度；当CaCl2的浓度达到25mg/L时，Kc形成复层，集落中央出现大量分化程度较高的角化细胞和角化物，周边仍存在增殖能力较强的单层Kc，逐渐向四周扩展。随着Ca2＋浓度的增高，细胞角化程度也随之增高。结论  Ca2+浓度对体外培养的Kc贴壁、增殖、分化等不同阶段产生影响，通过控制无血清培养液中Ca2+浓度可以调控Kc的增殖和分化。

    【关键词】  钙离子  角质形成细胞  分化  人工皮肤  组织工程

     Calcium is one of the key factors adjusting proliferation and differentiation of epidermal keratinocytes cultured in vitro

        【Abstract】  Objective  To explore the effects of calcium at different levels on proliferation and differentiation of epidermal keratinocytes cultured in serum free medium in vitro'  which is the base of tissue engineering skin.
Methods   Second passage epidermal keratinocytes of neonatal rabbits were seeded into 96-poles plate according to the cell density of 0.5×104/cm2'and cultured with serum free medium contained different levels of CaCl2 for 1 week.The cultured keratinocytes were observed' and identified with immunohistochemical test. Then' MTT test was used to detect the rela

[1] [2] [3] 下一页