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Xiangdan Wang

Publications and source records attributed to Xiangdan Wang.

2 recordsLinked to original sources

Microdialysis sampling membrane performance during in vitro macromolecule collection.

Microdialysis sampling is well-established for sampling small molecules. Recently, there has been an increased interest toward collecting macromolecules using microdialysis sampling. In this work, fluorescein isothiocyanate-labeled dextrans (FITC-dextrans) with molecular weight between 10 and 70 kDa were chosen as representative molecules to study analyte mass transport properties during microdialysis sampling using different lengths (2 and 10 mm) of 100-kDa MWCO polyethersulfone membranes. Experiments were performed in both well-stirred and quiescent phosphate-buffered saline solutions as well as in a 0.3% agar solution. Different fundamental parameters affecting microdialysis sampling of macromolecules, including effective membrane diffusion coefficients, were evaluated. The applicability of the most-often-cited Bungay et al. mass transfer model was compared to experimental data for the FITC-dextrans. For the larger macromolecules, the membrane provides a significant mass transport resistance most likely caused by hindered diffusion. These experimental aspects that are critical to microdialysis sampling of macromolecules are presented.

Dextrans↗

Multiplexed cytokine detection in microliter microdialysis samples obtained from activated cultured macrophages.

Microdialysis sampling probes were used to collect cytokine samples from lipopolysaccharide (LPS)-stimulated macrophages. The probes were immersed into cell culture wells containing either RAW 264.7 or isolated peritoneal macrophages. Dialysates (15 microL) from these wells were subjected to a multiplexed cytokine sandwich immunoassay platform analyzed by flow cytometry that measures up to six separate cytokines, interleukin-6 (IL-6), interleukin-10 (IL-10), interleukin-12p70 (IL-12p70), interferon-gamma (IFN-gamma), macrophage chemoattractant protein-1 (MCP-1), and tumor necrosis factor-alpha (TNF-alpha) in a single 15-muL sample. In vitro microdialysis sampling relative recovery experiments showed that only IFN-gamma, IL-6, MCP-1, and TNF-alpha could be recovered across a commercially-available 100-kDa MWCO microdialysis membrane. Eleven hours after LPS addition (1 microg/mL), RAW 264.7 macrophages secreted greater than 8000 pg/mL of TNF-alpha and greater than 1000 pg/mL MCP-1. With the peritoneal macrophages, greater than 6000 pg/mL of IL-6, MCP-1, and TNF-alpha were obtained. The maximum dialysate concentrations obtained from the RAW macrophages were 1300 pg/mL for TNF-alpha and 55 pg/mL for MCP-1. Maximum cytokine concentrations from peritoneal macrophage dialysates reached approximately 2000 pg/mL, 1100 pg/mL and 500 pg/mL for TNF-alpha, MCP-1 and IL-6, respectively. Microdialysis sampling allowed for 20-min samples to be collected during the cytokine release from the activated macrophages. These results demonstrate that microdialysis sampling can be used for collection of selected cytokines with improved temporal resolution.

Animals↗