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1.
Biotechnol Bioeng ; 33(10): 1235-41, 1989 Apr 20.
Article in English | MEDLINE | ID: mdl-18587855

ABSTRACT

Two human squamous carcinoma cell lines and human diploid fibroblasts were examined for the production of extracellular matrix (ECM) molecules including fibronectin (FN), laminin (LN), and thrombospondin (TSP) when grown on a number of different substrates. The substrates used included glass, plastic, collagen (gelatin), and DEAE-dextran. Levels of TSP as indicated by enzyme-linked immunosorbent assay did not vary significantly as a function of substrate. In contrast, LN levels in the culture medium were significantly decreased when the cells were grown on DEAE-dextran or collagen-linked dextran as compared to the other substrates. FN levels were slightly lower in the culture medium of the cells grown on DEAE-dextran. Biosynthetic labeling followed by immunoprecipitation indicated that the reduction in LN was due, in part, to decreased biosynthesis. Previous studies have indicated that LN influences the behavior of epithelial cells in culture and that the cells, themselves, are a major source of the LN. The differences in LN production noted here indicate that the production of this ECM component is influenced by the substratum on which the cells are grown. These differences could contribute to alterations in biological properties that are known to be influenced by the substratum.

2.
J Biol Stand ; 16(4): 333-8, 1988 Oct.
Article in English | MEDLINE | ID: mdl-2848844

ABSTRACT

Herpes simplex virus type I (HSV-1), infectious bovine rhinotracheitis virus (IBR) and turkey herpesvirus were examined for growth in cells cultured on three different substrates. The substrates were glass, DEAE-dextran and collagen gel. With two of the viruses, HSV-1 and IBR, there were no apparent differences in production as a function of substrate. In contrast, the amount of the turkey herpesvirus which was recovered varied greatly with the substrate. Titers were highest on glass, followed by DEAE-dextran and then collagen gel. Our previous studies have indicated that the substrate on which anchorage-dependent cells are grown in vitro has an affect on a number of biological and biochemical properties. The present study indicates that the production of commercially important biologicals can be affected by the substrate.


Subject(s)
Virus Cultivation/instrumentation , Animals , Cells, Cultured , Collagen , DEAE-Dextran , Glass , Herpesviridae/growth & development , Herpesvirus 1, Bovine/growth & development , Humans , Simplexvirus/growth & development , Surface Properties
3.
Lab Invest ; 59(2): 292-5, 1988 Aug.
Article in English | MEDLINE | ID: mdl-3261375

ABSTRACT

Tumor necrosis factor has a variety of effects on different types of cells in vitro, including endothelial cells. The current studies show that pretreatment of rat pulmonary artery endothelial cells with tumor necrosis factor increases in a time- and dose-dependent manner their sensitivity to killing by neutrophils stimulated with phorbol myristate acetate or C5a. Similar effects are seen with interleukin-1. These data suggest that tumor necrosis factor and interleukin-1 may have important pro-inflammatory effects on endothelial cells.


Subject(s)
Endothelium, Vascular/cytology , Neutrophils/immunology , Tumor Necrosis Factor-alpha/pharmacology , Animals , Cell Survival , Cells, Cultured , Complement C5/pharmacology , Dose-Response Relationship, Drug , Endothelium, Vascular/immunology , Humans , Interleukin-1/pharmacology , Rats , Tetradecanoylphorbol Acetate/pharmacology
4.
J Biol Stand ; 14(4): 331-6, 1986 Oct.
Article in English | MEDLINE | ID: mdl-3558416

ABSTRACT

Three commercially-important types of cell were grown on four different microcarrier substrates. The cells, which included normal human diploid fibroblasts (MRC-5), primary chick embryo cells and Madin-Darby bovine kidney cells (MDBK), were compared with regard to proliferation on the substrates and with regard to recovery of viable cells from the same substrates. The substrates used included glass-coated microcarriers (Biosil), collagen microcarriers (Ventregel), DEAE-dextran microcarriers (Cytodex I) and collagen-linked DEAE-dextran microcarriers (Cytodex III). The established cell line (MDBK) grew well on all of the substrates and a high percentage of viable cells could be harvested from each substrate. The MRC-5 cells also grew well on all four substrates but high recovery rates were achieved only with cells grown on the glass-coated microcarriers or collagen microcarriers. In contrast, the primary chick embryo cells grew well only on the glass microcarriers and the recovery rate of cells harvested from this substrate was high. In some industrial operations, the re-utilization of cells after removal from the substrate is necessary. In these situations the appropriate choice of microcarriers for the cultivation of the cells may be critical.


Subject(s)
Cell Division , Culture Media/analysis , Cell Adhesion/drug effects , Cell Count , Cell Line , DEAE-Dextran , Extracellular Matrix/physiology , Fibroblasts/physiology , Glass , Humans , Trypsin/pharmacology
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