ABSTRACT
The traditional primary culture methods of human normal epithelial cells have disadvantages of low activity of cultured cells,the low cultivated rate and complicated operation.To solve these problems,researchers made many studies on culture process of human normal primary epithelial cell.In this paper,we mainly introduce some methods used in separation and purification of human normal epithelial cells,such as tissue separation method,enzyme digestion separation method,mechanical brushing method,red blood cell lysis method,percoll layered medium density gradient separation method.We also review some methods used in the culture and subculture,including serum-free medium combined with low mass fraction serum culture method,mouse tail collagen coating method,and glass culture bottle combined with plastic culture dish culture method.The biological characteristics of human normal epithelial cells,the methods of immunocytochemical staining,trypan blue exclusion are described.Moreover,the factors affecting the aseptic operation,the conditions of the extracellular environment,the conditions of the extracellular environment during culture,the number of differential adhesion,and the selection and dosage of additives are summarized.