Cultured In Vitro And Histological Structure Study Of Human Epidermal Sheet

Cultured In Vitro and Histological structure study of Human Epidermal sheetObjective:To improve the techniques of human keratinocytes cultured in vitro, including the isolation, culture system and proper seeding concertration about human keratinocytes,in order to form HES that can be used to graft clinically.The micro and supermicro structure of HES were detected too.Methods: 1 Different methods were chosen for keratinocytes isolation and different culture conditions were used to culture keratinocytes in vitro.2 The relationship between seeding concentration and time of confluence was investigated by seeding keratinocytes at different concentrations.3 The HES was observed and identified by general,microscopical TEM and immunohistochemical methods.Results:1 The isolation methods of using Dispasell and Trypsin/EDTA were ideal,and the culture system containing 10%FCS, 3T3 feeder layer was better. 2 The proper seeding concentration of human keratinocytes was 2+106/75cm2 flask. 3 HES cultured in vitro was basically considered suitable for clinical application.Conclusion:Normal human keratinocytes that are isolated by using Dispase II and Trypsin/EDTA were able to survive, proliferate and confluent to form HES in culture system containing 10%FCS and 3T3 feeder layer at seeding concentration of 2+106/75cm2flask in 7-9 days.The HES was integrated and tight connection in histological examine,it's flexible and can be took off from flask and suit for graft.