A cobblestone cell isolated from the human omentum: the mesothelial cell; isolation, identification, and growth characteristics.
Normal human mesothelial cells (NHMC) were isolated from pieces of human omentum. The cell yield was approximately one million cells per square centimeter omentum. The mesothelial cells were identified by their positive staining with monoclonal antibodies against cytokeratins 6 and 18. Transmission electronmicroscopy of cultured NHMC revealed many microvilli on the apical surface and many mitochondria and pinocytotic vesicles in the cytoplasm, indicating active transmembrane transport. Growth of NHMC was directly related to the concentration of human serum or of fetal bovine serum in the growth medium. Addition of epidermal growth factor with or without hydrocortisone resulted in a significant increase of NHMC growth; when endothelial cell growth factor, insulin, or hydrocortisone were added no such increase was observed. Seeding NHMC at densities less than 3000/cm2 did not result in monolayer formation. The mesothelial cells were serially passed in growth medium M199 with added 10% fetal bovine serum up to 7 passages. However, after Passage 4 the cells changed into giant cells with an irregular pattern, and a lack of intracellular cytokeratin expression was observed for most of the cells.
Pronk
A
A
Laboratory of Surgery, University Hospital Utrecht, The Netherlands.
Leguit
P
P
Hoynck van Papendrecht
A A
AA
Hagelen
E
E
van Vroonhoven
T J
TJ
Verbrugh
H A
HA
D000911:Antibodies, Monoclonal; D002448:Cell Adhesion; D002455:Cell Division / Q000187:drug effects; D002469:Cell Separation / Q000379:methods; D003470:Culture Media; D046508:Culture Techniques / Q000379:methods; D016228:Endothelial Growth Factors / Q000494:pharmacology; D004815:Epidermal Growth Factor / Q000494:pharmacology; D004847:Epithelial Cells; D004848:Epithelium / Q000187:drug effects; D006133:Growth Substances / Q000494:pharmacology*; D006801:Humans; D006854:Hydrocortisone / Q000494:pharmacology; D007150:Immunohistochemistry; D007328:Insulin / Q000494:pharmacology; D007633:Keratins / Q000032:analysis; D007700:Kinetics; D009852:Omentum; D013997:Time Factors
D000911:Antibodies, Monoclonal; D003470:Culture Media; D016228:Endothelial Growth Factors; D006133:Growth Substances; D007328:Insulin; D004815:Epidermal Growth Factor; D007633:Keratins; D006854:Hydrocortisone