摘要
目的:探索和寻找人角朊细胞体外培养适宜的滋养层细胞,完善人角朊细胞体外培养快速扩增的技术和方法。方法:实验于2003-08/2004-03在武装警察部队总医院烧伤整形科组织工程实验室完成。实验分别在3T3细胞、原代培养的人成纤维细胞上接种原代人角朊细胞,并设无滋养层细胞的空白对照。在对不同滋养层细胞的处理上分别采用了60Co放射线照射的传统方法和丝裂霉素处理,比较两种处理方法对滋养层细胞的影响,并对丝裂霉素适宜的作用浓度进行了研究。结果:①丝裂霉素和60Co放射线照射分别处理滋养层细胞,可取得相同效果。②终浓度为15mg/L的丝裂霉素C是制备人成纤维细胞滋养层的较适宜浓度;终浓度为10mg/L的丝裂霉素C是制备3T3细胞滋养层的较适宜浓度。③原代人角朊细胞在不同滋养层细胞上的生长速度显示:人成纤维细胞>3T3细胞>无滋养层细胞。结论:经丝裂霉素处理的原代培养人成纤维细胞作为滋养层细胞,体外扩增人角朊细胞可以缩短培养时间,减少接种密度。
AIM:To search and seek for trophoblastic cells suitable for culture of human keratinocytes in vitro,and perfect a procedure for rapid amplification of human keratinocytes in vitro. METHODS:The experiment was conducted in the Laboratory of Tissue Engineering,Department of Burn and Plastic Surgery,General Hospital of Chinese People's Armed Police Forces during August 2003 and March 2004.Primary human keratinocytes were inoculated into 3T3 cells or primary cultured human fibroblasts as trophoblastic cells, and the cells without feeder layer were used as controls.60Co radiation and mitomycin of different concentration were used to manage different trophoblastic cells,and the effects of the two treatments on the trophoblastic cells were compared.The suitable concentration of mitomycin was also studied. RESULTS:①The trophoblastic cells treated with mitomycin had the same function as those treated with 60Co radiation.②The final concentration of 15 mg/L was suitable for mitomycin to prepare the feeder layers of human fibroblasts,and 10 mg/L suitable for mitomycin to prepare 3T3 cells.③The proliferation speed of primary human keratinocytes on different trophoblastic cells was in the following sequence:human fibroblasts >3T3 cells >cells without feeder layers. CONCLUSION:Primary cultured human fibroblasts treated with mitomycin can be served as trophoblastic cells.Amplification of human keratinocytes in vitro can shorten culture time and decrease inoculation density.
出处
《中国临床康复》
CSCD
2004年第35期7945-7947,i001,共4页
Chinese Journal of Clinical Rehabilitation