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You are here: Home / Associated publications / Belgian Journal of Zoology / Bibliographic References / Method for culture of a bovine pulmonary endothelial cell strain (CPAE) in a serum-free medium and on microporous membranes coated with matrigel ™.

S. Baatout, F. Kinard, Claude Remacle and Y-J. Schneider (1995)

Method for culture of a bovine pulmonary endothelial cell strain (CPAE) in a serum-free medium and on microporous membranes coated with matrigel ™.

BELGIAN JOURNAL OF ZOOLOGY, 125(1):53-66.

In order to optimize culture conditions of bovine pulmonary endothelial cells (CPAE), we have compared different culture media, supports and extracellular matrices. Cell biomass was estimated by protein assay using Lowry's method. Different constituents including albumin, hydrocortisone, insulin, transferrin, triiodotyronine, Epidermal Growth Factor (EGF) and Fibroblast Growth Factor were separately added to Basal Defined Medium (BDM). Among them, BDM supplemented with EGF (1 ng/ml), hydrocortisone (100 nM), insulin (1 mu g/ml), triiodotyronine (2 nM) and linoleic acid complexed to albumin (10 mu g/ml) also named `synthetic BDM' appeared to be the best serum-free nutritive medium and showed similar results as compared to DMEM supplemented with 10\% Fetal Bovine Serum (FBS). Several supports have then been tested including tissue culture polystyrene (as a reference), teflon, polycarbonate or poly(ethylene terephthalate) track-etched membranes. Among them, cells cultivated on surface treated membranes in poly(ethylene terephthalate) exhibited the highest protein content with a significant increase in comparison to tissue culture polystyrene, probably because cells are fed on the two faces instead of one. On treated poly(ethylene terephthalate) membranes, cells kept their endothelial morphology and ultrastructure. Finally, cell biomass on several exogenous extracellular matrices was studied. Cells were cultivated in `synthetic BDM' or DMEM supplemented with 10\% FBS and on poly(ethylene terephthalate) membranes. Among fibronectin, matrigel(TM) (solubilized tissue basement membrane), laminin, collagen (type I) and polylysine; matrigel(TM) appeared to be the optimal extracellular matrix.

CPAE CELLS; IN VITRO MODEL; SERUM-FREE MEDIUM; MICROPOROUS MEMBRANES; POLY(ETHYLENE TEREPHTHALATE); EXTRACELLULAR MATRIX; MATRIGEL
1st Benelux Congress on Zoology, LEUVEN, BELGIUM, NOV 04-05, 1994
BJZ

ISSN 2295-0451 (online version)
ISSN 0777-6279 (printed version)
impact factor 2015: 0,87.

Editor-in-Chief:
Prof. Dr. Isa Schön
Royal Belgian Institute of Natural Sciences
Vautierstraat 29
1000 Brussels, Belgium

 



1863-1903
Annales de la Société malacologique de Belgique
 
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Belgian Journal of Zoology