Anchorage-independent growth of normal human mesothelial cells: A sensitive bioassay for EGF which discloses the absence of this factor in fetal calf serum |
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Authors: | Paul J La Rocca James G Rheinwald |
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Institution: | (1) Division of Cell Growth and Regulation, Dana-Farber Cancer Institute, Harvard Medical School, 44 Binney Street, 02115 Boston, MA;(2) Department of Physiology and Biophysics, Harvard Medical School, 44 Binney Street, 02115 Boston, MA |
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Abstract: | Summary This laboratory recently reported that normal human mesothelial cells require epidermal growth factor (EGF) and hydrocortisone
(HC), in addition to fetal calf serum and a complex defined medium component, in order to grow optimally in surface culture
(9). We report here that this normal cell type also forms large colonies at high efficiency in semi-solid medium, but exhihits
more stringent serum and EGF requirements for anchorage-independent than for surface growth. Mesothelial cells are unable
to divide at all in semi-solid medium with added EGF or with less than 2% serum, whereas they grow slowly but progressively
in surface culture under such conditions. In semi-solid medium containing 20% serum and HC, mesothelial cells are stimulated
to divide by the addition of as little as 30 pg/ml purified EGF. Human urine or male mouse plasma could substitute for purified
EGF, yielding growth commensurate with the levels of EGF in these biological fluids previously measured by others using radioreceptor
and radioimmune assays. Thus growth of mesothelial cells in semi-solid medium can serve as a highly sensitive assay of EGF
biological activity which is unaffected by the presence of serum proteins. In addition, our results demonstrate that fetal
calf serum does not provide mitogenic levels of EGF to cultured cells, raising the question of the identity of plasma and
serum mitogens.
This work was supported by NIH grants RO1 AG02048 and RO1 CA26656 to James G. Rheinwald and by NIH postdoctoral fellowship
F32 AG05303 to Paul J. La Rocca. |
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Keywords: | EGF mesothelial cells anchorage-independent growth human diploid |
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