A cobblestone cell isolated from the human omentum: the mesothelial cell; isolation,identification, and growth characteristics |
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Authors: | Apollo Pronk Piet Leguit Arthur A G M Hoynck van Papendrecht Edwin Hagelen Theo J M V van Vroonhoven Henri A Verbrugh |
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Institution: | (1) Laboratory of Surgery, University Hospital Utrecht, Utrecht, The Netherlands;(2) Laboratory of Microbiology, University Hospital Utrecht, Utrecht, The Netherlands;(3) Diakonessenhuis and the Laboratory of Surgery, University Hospital Utrecht, Utrecht, The Netherlands;(4) Department of Surgery, University Hospital Utrecht, 3808 GA, Post Box 85500, Utrecht, The Netherlands |
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Abstract: | Summary 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. |
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Keywords: | mesothelial cells mesothelial cell identification peptide growth factors serial propagation |
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