Primary culture of human esophageal epithelial cells

Abstract

Primary cultures and serial passages of nontumorous adult human esophageal epithelial cells derived from surgically resected esophagi of patients with esophageal cancer have been developed. Disaggregated cells obtained by means of tryptic digestion lasting for 90 min grew to confluency in medium RITC 80-7 supplemented with 10% fetal bovine serum (FBS). Such primary cultures were established from more than 20 donors, and could be passaged 3 to 5 times. Fibroblastic contamination was almost eliminated by careful sampling, and it was possible to perform the growth response or serial passages without fibroblastic overgrowth. Serum-free medium RITC 80-7 supplemented with fibronectin (FN) and bovine serum albumin (BSA) also supported the growth of esophageal epithelial cells, but was not sufficient for serial passages. Among growth promoting factors in medium RITC 80-7, the epidermal growth factor (EGF) was highly stimulatory to growth. Insulin, transferrin, FN and BSA also enhanced cell growth and/or attachment. The epithelial nature of the cultured cells was confirmed by transmission electron microscopy and indirect immunofluorescent staining with antikeratin antibody. The majority of the cultured cells were diploid.