GTSF-2: A new, versatile cell culture medium for diverse normal and transformed mammalian cells

Abstract

The aim of this study was to test the versatility of a new basal cell culture medium, GTSF-2. In addition to traditional growth-factors, GTSF-2 contains a blend of three sugars (glucose, galactose, and fructose) at their physiological levels. For these studies, we isolated normal endothelial cells from human, bovine, and rat large blood vessels and microvessels. In addition, GTSF-2 was also tested as a replacement for high-glucose-containing medium for PC12 pheochromocytoma cells and for other, transformed cell lines. The cell growth characteristics were assessed with a novel cell viability and proliferation assay, which is based on the bioreduction of the fluorescent dye, Alamar Blue. After appropriate calibration, the Alamar Blue assay was found to be equivalent to established cell proliferation assays. Alamar Blue offers the advantage that cell proliferation can be measured in the same wells over an extended period of time. For some of the cell types (e.g., endothelial cells isolated from the bovine aorta, the rat adrenal medulla, or the transformed cells), proliferation in unmodified GTSF-2 was equivalent to that in the original culture media. For others cell types (e.g., human umbilical vein endothelial cells and PC12 cells), GTSF-2 proved to be a superior growth medium, when supplemented with simple additives, such as endothelial cell growth supplement (bFGF) or horse serum. Our results suggest that GTSF-2 is a versatile basal medium that will be useful for studying organ-specific differentiation in heterotypic coculture studies.

A549 Lung Epithelial Cells Grown as Three-Dimensional Aggregates: Alternative Tissue Culture Model forPseudomonas aeruginosaPathogenesis

“…The human tumorigenic lung epithelial cell line A549 was obtained from the American Type Culture Collection (CCL-185). A549 cells to be cultured in three dimensions were initially grown in Corning T75 cell culture flasks containing GTSF-2 (HyClone) (22) supplemented with 10% fetal bovine serum and 100 g of penicillin-streptomycin (Invitrogen) per ml at 37°C in 5% CO 2 before being seeded into the RWV bioreactor. On reaching 75% confluency, the A549 cells were washed with phosphate-buffered saline (PBS) and then removed from the flask by the addition of 0.25% trypsin and washed twice with PBS.…”

Vesicle traffic through intercellular bridges in DU 145 human prostate cancer cells

“…Human prostate cancer cells DU 145 (HTB 81) were purchased from American Type Culture Collection (Manassas, VA), and cultures were propagated at 37ºC, 5% CO 2 and 95% relative humidity in GTSF-2 complete medium [13] containing 7% fetal bovine serum (pH 7.4). Stock cultures of this cell line were maintained in T-flask and subcultured by trypsinization.…”

Characterization of the Contribution to Virulence of Three Large Plasmids of Avian Pathogenic Escherichia coli χ7122 (O78:K80:H9)

“…The cell culture media used included Dulbecco's modified Eagle's medium (DMEM) (Gibco), RPMI-1640 (Gibco), and GTSF-2 medium consisting of triple-sugar minimal essential medium ␣-L-15 base supplemented with 2.2 g/liter NaHCO 3 and 2.5 mg/liter insulin-transferrin-sodium selenite (40).…”

“…At different steps of infection, the pathogen has to acquire nutrients that are necessary for its growth and survival. In this study, we evaluated the in vitro growth of bacteria in both bacterial culture media and cell culture media that are designed to maintain cells under the conditions found in the original tissue (40). The results of this study showed that plasmid-containing derivatives of APEC 7122 grow differently in the different media tested.…”