人皮肤角质形成细胞的制备及体外培养

目的 探讨人皮肤角质形成细胞制备及体外培养的方法.方法 皮肤标本用胰酶、胰酶＋分离酶（dispase酶）2种方法消化,并分别以DMEM培养基、RPMI 1640培养基、限制性角质形成细胞无血清培养基（KC-FSM培养基）培养,MTT法通过比色分析测定培养48 h、72 h后细胞的光密度值（D值）,以判断不同培养基对角质形成细胞增殖的影响.结果 胰酶＋分离酶消化可获得较纯的原代角质形成细胞,细胞在限制性KC-FSM培养基中生长良好,培养48 h、72 h后测得的D值显著高于DMEM培养基和RPMI 1640培养基（P＜0.05）.结论 胰酶＋分离酶消化、限制性KC-FSM培养基体外培养是制备和培养人皮肤角质形成细胞的好方法.

Preparation and culture of human skin keratinocytes in vitro

Objective To investigate the approaches to the preparation and culture of human skin keratinocytes in vitro. Methods Two methods were used to digest human skin specimen was by trypsogen as well as trypsogen and dis- pose, followed by culture in DMEM medium, RPMI 1640 medium and defined KC - FSM medium, respectively. The optical density （D） values of cells at 48 h and 72 h of incubation were determined by colorimetric assay of MTF to evaluate the effects of different culture media on the proliferation of keratinocytes. Results Comparatively pure primary keratinocytes could be obtained if the skin specimen was digested by the use of trypsogen plus dispose enzyme. Cell proliferation of human skin keratinocytes was satisfactory and D value was significantly higher if cells were incubated with defined KC - FSM medium for 48 h and 72 h （P 〈 0.05）. Conclusion Trypsogen plus dispose enzyme and defined KC - FSM medium proves to be good methods for the preparation and culture of human skin keratinocytes in vitro.