|| Caputo J.L., Hay R.J., Williams C.D.|
||The isolation and properties of an epithelial cell strain from rhesus monkey bronchus.|
|| In Vitro 15:222-223(1979)|
||An epithelial cell strain dependent upon epidermal growth factor (EGF) for
proliferation has been isolated from explant cultures of monkey bronchi.
Segments of approximately 3 mm2 were placed in 60 mm plates containing
glass chips. The standard growth medium used was F12K with 15% FBS. Chips
supporting epithelial outgrowths apparently free of fibroblasts were
transferred to 16 mm wells of cluster plates. Cells thus isolated required
both EGF (10-30 ng/ml) and the presence of irradiated lung fibroblasts
(ATCC CCL-171 - MRC5) for subsequent propagation to mass population
densities. Epithelial cultures could be propagated serially for more than
6 months with over 33 population doublings. The omission of EGF at any
point resulted in an obvious decline in proliferation rates and the
appearance of vacuolated cells. Biochemical studies showed that addition
of EGF resulted in a 20-fold increase in 3H-thymidine incorporation.
Dexamethasone (10^-8 M) acted synergistically with EGF to double
3H-thymidine uptake. Histochemical staining showed that vigorous cultures
contained abundant PAS-positive cytoplasmic droplets. The cells also
exhibited high levels of malic dehydrogenase as did epithelia lining
bronchial lumina. Examination of the fine structure of cultured bronchial
cells revealed desmosomes plus a variety of electronlucent droplets and
dense membrane-bound bodies with somewhat heterogeneous matrices. The cell
strain has been preserved in liquid nitrogen and can be recovered readily.
Further characterizations are in progress.
|| CVCL_4492; 4MBr-5|