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A flow bioreactor system compatible with real-time two-photon fluorescence lifetime imaging microscopy.

Nian Shen1, Julia A Riedl, Daniel A Carvajal Berrio

  • 1Department of Women's Health, Research Institute of Women's Health, University Hospital of the Eberhard Karls University, Tübingen, Germany. Department of Cell and Tissue Engineering, Fraunhofer Institute for Interfacial Engineering and Biotechnology IGB, Stuttgart, Germany.

Biomedical Materials (Bristol, England)
|November 18, 2017
PubMed
Summary

This study introduces a novel bioreactor for continuous cell culture analysis, enabling real-time monitoring of cell behavior and metabolism under fluid shear stress. The system allows for non-invasive, high-resolution imaging, overcoming limitations of traditional methods.

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Area of Science:

  • Biomedical Engineering
  • Cell Biology
  • Microfluidics

Background:

  • Bioreactors are crucial for in vitro cell and tissue culture.
  • Current bioreactors require experiment interruption for sample analysis.
  • A need exists for continuous, non-invasive monitoring in bioreactor systems.

Purpose of the Study:

  • To design and validate a novel bioreactor system for continuous, high-resolution imaging of cells under flow.
  • To investigate the effects of shear stress on human umbilical vein endothelial cells (HUVECs).
  • To enable real-time monitoring of cellular responses and metabolic shifts.

Main Methods:

  • Development of a closed-system bioreactor with continuous flow and microscope compatibility.
  • Integration with two-photon fluorescence lifetime imaging microscopy (2P-FLIM).
  • Hydrodynamic characterization using COMSOL simulations and shear stress analysis.
  • In vitro experiments on HUVECs, including scratch assays and metabolic activity measurements.

Main Results:

  • The bioreactor demonstrated homogeneous and reproducible flow conditions.
  • Low shear stress significantly decreased HUVEC migration in a scratch assay.
  • 2P-FLIM detected a metabolic shift from glycolysis to oxidative phosphorylation in HUVECs under shear stress.

Conclusions:

  • The developed bioreactor facilitates real-time, contact-free imaging and analysis of cellular responses to biophysical stimuli.
  • This system provides novel insights into cell migration and metabolic changes under shear stress.
  • Potential applications include real-time cell fate tracking and drug efficacy monitoring.