人的原代上皮角质形成细胞和永生化的上皮角质形成细胞HaCaT的增殖能力的比较

Cell Prolifereration Capacity of Comparation Keratinocytes and Immortalization Epidermal HaCaT

目的:筛选适合充当组织工程皮肤的种子细胞,比较原代培养的人原代上皮角质形成细胞和永生化的上皮角质形成细胞HaCaT的增值能力。方法:将两种细胞(小儿包皮环切术后的组织培养表皮角质形成细胞,永生化的上皮角质形成细胞HaCaT),分别接种于96孔板,通过MTT检测细胞1,3,5,7,9,11天的生长情况;当两种细胞融合至60%时分别取1×106个细胞,通过流式细胞检测细胞的周期。结果:永生化的上皮角质形成细胞HaCaT每隔一天即可传代一次,原代上皮角质形成细胞每3天传代一次;细胞周期:永生化上皮角质形成细胞HaCaT的G1期和S期的比例高于原代上皮角质形成细胞。生长曲线:MTT检测两种细胞1,3,5,7,9,11的生长情况,永生化的上皮角质形成细胞HaCaT的生长速度明显高于原代上皮角质形成细胞。结论:永生化上皮角质形成细胞HaCaT的增殖能力要高于原代培养的上皮角质形成细胞。

Objective： To compare proliferation capacity of human primary keratinocytes and immortalized epiderm cell line HaCaT.Methods： Getting normal skin biopsy samples（sampleswere obtained from foreskin circumcision） to culture keratinocytes,immortalized epidrem cell line Hacat,two kinds of cells were seeded in 96 well plates,cells growth curve were detected by MTT,when two kinds cells mixed 60% taking 1 × 106 cells by flow cytometry to detect cell cycle.Results： Immortalized epiderm cell line Hacat can be passaged once every other day,human primary keratinocytes can be passaged every three days.Cell cycle： immortalized epiderm HaCat G1 phase and S phase was higher than primary keratinocytes.Growth curve： MTT detected two kinds cells 1,3,5,7,9,11 growth situation,immortalized epiderm cell line HaCat grow faster than primary keratinocytes.Conclusion： Immortalized epiderm HaCat proliferation ability better than primary keratinocytes.