from BD Biosciences - Discovery Labware

The purpose of this study was to evaluate the effect of BD Matrigel™ Matrix Thin-Coat and BD Matrigel™ Matrix Overlay on the attachment, morphology, and CYP enzyme activities of human cryopreserved hepatocytes. BD Gentest™ Human CryoHepatocytes were tested for basal and induced CYP3A4 and CYP1A2 activities on Collagen I and BD Matrigel Matrix thin-coat surface with or without a BD Matrigel Matrix overlay. Changes in attachment and morphology were also examined. The results showed that the BD Matrigel Matrix thin-coat surface significantly improved basal activities for both CYP3A4 and CYP1A2 compared to a conventional collagen I-coated surface; the BD Matrigel Matrix thin-coat surface showed a lot-dependent effect towards induced activities for both CYP3A4 and CYP1A2 for tested lots. The BD Matrigel Matrix overlay, in addition to the BD Matrigel Matrix thin-coat plate, further improved basal CYP450 activities, however, the BD Matrigel Matrix overlay did not change cell attachment.The BD Matrigel Matrix thin-coat can maintain typical hepatocyte morphology for a longer time than the collagen I-coated surface. In conclusion, a culture condition combining BD Matrigel Matrix thin-coat surface and BD Matrigel Matrix overlay has a potential of maintaining stable long-term basal metabolic activities for human cryopreserved hepatocytes, facilitating their application in areas such as in vitro chronic toxicity assays. Read More

The interaction between extracellular matrix (ECM) and neural cells is important in cell differentiation and morphogenesis of the nervous system. However, conditions for culture of neuronal cells have not been optimized for ECM, and poly-lysine often prevails as the substrate of choice. Quantitative Ca⁺⁺ fluorimetry was used to screen ECM proteins singly and in combination to determine culture conditions resulting in optimal morphology and function of rat cortical cells. A fluid handling robot was used to ensure uniformity in cell plating and reagent dispensing. Agonist responses in cells from single isolations plated on laminin followed by human fibronectin (LM/HFN) were significantly enhanced when compared with cells cultured on LM or HFN alone, as well as on poly-lysine or uncoated tissue culture plastic. In addition, differences in morphology between cultures grown on the various substrates were observed. Pyramidal neurons exhibited long interconnecting processes with growth cones on LM/HFN. Neurites in LM or HFN cultures were significantly shorter and cell bodies formed aggregates on LM. Elongated neurites were characteristic of cells grown on poly-lysine. Preliminary studies using time lapse confocal microscopy with a fluorescent Ca⁺⁺ dye (Fluo3AM) indicated that glutamate receptor function also varies in cells cultured on different ECM substrates, correlating with immunological quantitation of the glutamate receptor (GluR1). Neural cells grown on LM/HFN displayed improved cell morphology and receptor function. These studies indicate that ECM is an important component to consider in obtaining optimal neural cell cultures. Read More

It's 5pm; do you know how clean your eukaryotic cell cultures are? They may not be as pristine as you think. Your culture dishes could be infected ... read more