A portion of the GFP-labled or unlabeled UCB MSCs (umbilical cord blood mesenchymal stem / stromal cells) were incubated with trypan blue and counted using a Cellometer Auto T4 cell counter (Nexcelom Bioscience LLC, Lawrence,MA) to determine the number of viable cells. […]
Leukocyte counts of BAL fluid BAL samples were mixed gently prior to diluting 50 ml BAL fluid with 50 ml of trypan blue (1:5 in 1xPBS) for viable cell counts. 20 ml of diluted cell sample were pipetted on a Cellometer counting chamber. Leukocytes were automatically counted using a Cellometer Auto T4 (Nexcelom Bioscience, Lawrence, MA, USA) […]
Viable T cells were enumerated every 7 days by trypan blue exclusion using the AutoT4 cell counter Cellometer (Nexcelom Bioscience, Lawrence, MA) […]
Canine adipose-derived mesenchymal stem cells (cASCs) When the monolayer of adherent cells reached approximately 70–80% confluence; trypsinization for cell splitting was performed using trypsin-EDTA solution (0.05%/0.02%, Biochrom AG, Berlin, Germany). After centrifugation, the cell pellets were subcultured at a concentration of 3 × 105cells/mL medium on 25cm2 tissue culture flasks containing DMEM/10% FCS. Cell counting was carried out by an automatic cell counter (Cellometer Nexcelom Bioscience, Lawrence, USA). The viable cell percentage from the harvested cells was calculated after trypan blue staining by an automatic cell counter (Cellometer Nexcelom Bioscience, Lawrence, USA). The cell concentration used for the proliferation assay [...]
Human non-small cell lung cancer (NSCLC) cell lines included the parental cell line A549 and the cell subline A549/DDP. Pt levels were expressed as μM/1 × 106 cells, with the cell number determined by counting (Nexcelom Cellometer Auto T4 cell counter, USA) in parallel cultures. Experiments were performed in duplicate and the values expressed were the means ± SD of the three independent experiments. […]
Gene expression actualizes the organismal phenotypes encoded within the genome in an environment-dependent manner. Among all encoded phenotypes, cell population growth rate (fitness) is perhaps the most important, since it determines how well-adapted a genotype is in various environments. Traditional biological measurement techniques have revealed the connection between the environment and fitness based on the gene expression mean. […]
Ficoll separation is routinely used to isolate mononuclear cells from bone marrow, peripheral blood, and umbilical cord blood. Monocytes and lymphocytes form a buffy coat under a layer of plasma, where the cells are collected, then washed. Typically, some residual platelets and red blood cells are mixed in with the mononuclear cells. The degree of platelet and red blood cell contamination can vary significantly between donors and operators. The Cellometer Auto 2000 Cell Viability Counter utilizes dual fluorescence counting to generate accurate live cell and dead cell counts with no interference from red blood cells and platelets. Experimental Procedure Combine 20µl [...]
Isolation, Quantitation and Viability Analysis of Neonatal Cardiomyocytes using Cellometer The neonatal rat cardiomyocyte model has been used for many years by researchers studying the heart. In addition to increasing understanding of the morphological, biochemical, and electrophysiological characteristics of the normal heart, neonatal cardiomyocytes have been used to study contraction, ischemia, hypoxia, and the toxicity of different compounds. This model has been used to determine the optimal dosage for certain drugs and to develop and evaluate the efficacy of potential therapeutic agents. Apoptosis, or programmed cell death, has been observed in infarcted and re-perfused myocardium, end stages of heart failure, [...]
Caspase-8 Caspase-8 belongs to a family of evolutionally conserved cysteine proteases that play a key role in regulating programmed cell death or apoptosis. The two main apoptosis activation pathways are the extrinsic and the intrinsic pathways. The extrinsic pathway is activated by the binding of ligands (including TNFα, FasL, and TRAIL) to cell-surface receptors and the conversion of procaspase-8 to the active caspase-8 protease.1 Caspase-8 quickly converts procaspase-3 to active Caspase-3, facilitating many of the cellular and biochemical events of apoptosis. The intrinsic, or mitochondrial, pathway is typically activated in response to DNA or cellular damage and begins with the [...]
Stem Cells are undifferentiated cells within a tissue or organ that can develop into specialized cell types. Embryonic Stem Cells are derived from human embryos developed from eggs fertilized in vitro and donated for research. Though stem cell research has shifted away from embryonic stem cells for ethical and legal reasons, there are currently 21 approved stem cell lines that are monitored by the NIH Stem Cell Unit. Undifferentiated cells derived from cultured embryonic stem cells, or pluripotent cell lines, can be used to test new drugs and could potentially be used in cell-based therapy for a wide range of [...]