Chemically induced growth inhibition and cell cycle perturbations in cultures of differentiating rodent embryonic cells

Cell Cycle Inhibition by Sodium Arsenite in Primary Embryonic Rat Midbrain Neuroepithelial Cells

“…Cell culture. Embryonic rat midbrain neuroepithelial cell cultures were prepared according to our previously established protocol (Flint, 1983;Ponce et al, 1994b;Ribeiro and Faustman, 1990;Faustman, 1991, 1992;Whittaker et al, 1993) with slight modifications. Gravid uteri were removed from pregnant (GD 12) Sprague-Dawley albino rats (Charles River, Hollester, CA) and placed in Earl's Balanced Salt Solution (EBSS, Gibco BRL, Grand Island, NY).…”

“…The present study examined the impact of concentrations (0.5-4 lM) of As 3þ on cell cycle kinetics of primary embryonic rat midbrain (GD 12) neuroepithelial cells. The embryonic midbrain neuroepithelial cell culture system has been used extensively by our group and others to examine the effects of toxicants on proliferation and differentiation of neuronal precursor cells on GD 12 (Flint, 1983;Ponce et al, 1994;Ribeiro and Faustman, 1990;Faustman, 1991, 1992;Whittaker et al, 1993). This culture system has been used to evaluate the effects of chemicals on cell viability, [ 3 H] c-aminobutyric acid uptake (George and Gabay, 1968), and hematoxylin-stained neurites (Flint and Orton, 1984).…”

“…The cells were non-immunoreactive for glial fibrillary acidic protein (GFAP) (Whittaker et al, 1993). This system has also been successfully utilized to characterize changes in cell cycling in response to exogenous factors such as alkylating agents, albendazole, methylmercury, chlorpyrifos, and serotonin (Cosenza and Bidanset, 1995;Menegola et al, 2004;Ponce et al, 1994;Ribeiro and Faustman, 1990;Seeley and Faustman, 1995;Whittaker and Faustman, 1991).…”

Micromass Cultures in Teratology

“…Although originally proposed as a screening tool for teratogens, the culture of midbrain neuroepithelial cells and limb bud chondrocytes has become important in vitro test models for examining the cellular and molecular processes governing normal development and the cellular response to environmental stresses that may influence development. For example, these culture systems have been used extensively by Faustman and colleagues to examine the mechanisms underlying the developmental toxicity of n ‐nitroso compounds, metals, and pharmaceutical and agricultural compounds (Ribeiro and Faustman, ; Whittaker and Faustman, , ; Sweeney et al, ; Whittaker et al, ; Faustman and Sweeney, ; Kidney and Faustman, ; Seeley and Faustman, ).…”

“…Because they are derived from primary cell suspensions of undifferentiated midbrain (mesencephalic) neuroepithelium and forelimb chondrocytes, micromass cultures have been used in the study of developmental cell cycle kinetics and regulation, which are poorly modeled by transformed cell lines. Micromass cultures have also been used to examine the cellular and molecular response to known teratogenic compounds, including analysis of protein and gene expression, intracellular cation signaling, redox status, necrosis and apoptosis, and differentiation (Flint et al, ; Brown et al, , ; Ribeiro and Faustman, ; Walum and Flint, ; Whittaker and Faustman, , ; Sweeney et al, ; Whittaker et al, ; Ponce et al, ; Seeley and Faustman, ; Ou et al, , ). As these studies demonstrate, micromass cultures can be successfully examined using a range of experimental tools, including flow cytometry and attached cell laser cytometry, gel electrophoresis, and light, electron, and immunofluorescence microscopy.…”