Deriving Quantitative Cell Biological Information from Dye-Dilution Lymphocyte Proliferation Experiments.
AuthorsRoy K, Shokhirev MN, Mitchell S, Hoffmann A,
JournalMethods Mol Biol
PubMed ID29388101
'The dye-dilution assay is a powerful tool to study lymphocyte expansion dynamics. By combining time course dye-dilution experiments with computational analysis, quantitative information about cell biological parameters, such as percentage of cells dividing, time of division, and time of death, can be produced. Here, we describe the method to generate ... More
Triple co-culture of human alveolar epithelium, endothelium and macrophages for studying the interaction of nanocarriers with the air-blood barrier.
AuthorsCosta A, de Souza Carvalho-Wodarz C, Seabra V, Sarmento B, Lehr CM,
JournalActa Biomater
PubMed ID31004840
'Predictive in vitro models are valuable alternatives to animal experiments for evaluating the transport of molecules and (nano)particles across biological barriers. In this work, an improved triple co-culture of air-blood barrier was set-up, being exclusively constituted by human cell lines that allowed to perform experiments at air-liquid interface. Epithelial NCI-H441 ... More
Isolation and Activation of Murine Lymphocytes.
AuthorsLim JF, Berger H, Su IH
JournalJ Vis Exp
PubMed ID27842342
'B and T cells, with their extremely diverse antigen-receptor repertoires, have the ability to mount specific immune responses against almost any invading pathogen(1,2). Understandably, such intricate abilities are controlled by a large number of molecules involved in various cellular processes to ensure timely and spatially regulated immune responses(3). Here, we ... More
Superior properties of CellTrace Yellow™ as a division tracking dye for human and murine lymphocytes.
The discovery of cell division tracking properties of 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester (CFSE) by Lyons and Parish in 1994 led to a broad range of new methods and numerous important biological discoveries. After labeling, CFSE is attached to free amine groups and intracellular proteins in the cytoplasm and nucleus of ... More
Autoreactive, Low-Affinity T Cells Preferentially Drive Differentiation of Short-Lived Memory B Cells at the Expense of Germinal Center Maintenance.
AuthorsJain RW, Parham KA, Tesfagiorgis Y, Craig HC, Romanchik E, Kerfoot SM,
JournalCell Rep
PubMed ID30566861
B cell fate decisions within a germinal center (GC) are critical to determining the outcome of the immune response to a given antigen. Here, we characterize GC kinetics and B cell fate choices in a response to the autoantigen myelin oligodendrocyte glycoprotein (MOG) and compare the response with a standard ... More
Colorectal tumor-on-a-chip system: A 3D tool for precision onco-nanomedicine.
Awareness that traditional two-dimensional (2D) in vitro and nonrepresentative animal models may not completely emulate the 3D hierarchical complexity of tissues and organs is on the rise. Therefore, posterior translation into successful clinical application is compromised. To address this dearth, on-chip biomimetic microenvironments powered by microfluidic technologies are being developed ... More