B16细胞与HaCaT细胞体外共培养模型的建立  

Construction of the co-culture model of B16 cells and HaCaT cells in vitro

作　　者：朱祯慧 朱丽清[1] 陈金妹[1] 林娇芬[1] 潘裕添[1] 

机构地区：[1]闽南师范大学菌物产业工程技术中心,福建漳州363000

出　　处：《陕西科技大学学报》2017年第4期132-137,共6页Journal of Shaanxi University of Science & Technology

基　　金：福建省科技厅青年科技创新项目(2015J05071)

摘　　要：初步建立小鼠黑色素瘤细胞(B16细胞)和人永生化角质形成细胞(HaCaT细胞)的共培养模型,利用熊果苷和8-甲氧补骨脂素(8-MOP)对此模型进行作用验证.使用10%DMEM完全培养基对B16细胞和HaCaT细胞进行培养,尝试不同的细胞接种顺序和细胞接种浓度构建共培养模型;将不同浓度的熊果苷和8-MOP作用到共培养模型中,通过测细胞毒性、黑色素含量和酪氨酸酶含量来验证模型的可靠性.结果表明,细胞接种顺序对共培养模型没有影响,采用B16细胞数量:HaCaT细胞数量为1∶4为最终细胞接种浓度比例;当熊果苷为100μg/mL时对黑色素含量和酪氨酸酶含量达到最高抑制效果;当8-MOP为20μg/mL时对黑色素含量和酪氨酸酶含量达到最高促进效果,均与单独B16细胞实验结果趋势一致.有效构建了B16细胞和HaCaT细胞共培养模型,并且可以将该模型运用于筛选抑制/促进黑色素药物中.The co-culture model of Murine Melanoma cells(B16 cells)and human keratinocytes(HaCaT cells)was established,and the effect of the model by arbutin and 8-MOP.B16 cells and HaCaT cells were cultured in 10% DMEM medium.Different inoculation sequences of cell and inoculation concentrations of cell were used to construct co-culture model.After arbutin and 8-MOP were added to the model,the cytotoxicity,melanin content and tyrosinase activity were measured to verify the reliability of the model.The results showed that the sequences of cell inoculation had no effect on the co-culture model;B16 cells was co-culture with HaCaT cells as 1∶4 ratio;the highest inhibitory effect of arbutin on the melanin content and tyrosinase activity at the concentration of 100 μg/ml;the 8-MOP had the lowest inhibitory effect on melanin content and tyrosinase activity at the concentration of 20μg/ml,all of these like as the previous study of only B16 cells.The co-culture model of B16 cells and HaCaT cells was established effectively in this study and it can be used to screen the drugs of promoting or inhibiting melanin.

关 键 词：B16细胞 HACAT细胞 共培养模型 熊果苷 8-MOP 

分 类 号：Q2-3[生物学—细胞生物学]