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Open Access Publications from the University of California

Noninvasive Quantification of Cell Density in Three-Dimensional Gels by MRI.

  • Author(s): Archer, Brian J
  • Uberruck, Till
  • Mack, Julia J
  • Youssef, Khalid
  • Jarenwattananon, Nanette N
  • Rall, Deniz
  • Wypysek, Denis
  • Wiese, Martin
  • Blumich, Bernhard
  • Wessling, Matthias
  • Iruela-Arispe, M Luisa
  • Bouchard, Louis-S
  • et al.

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OBJECTIVE:For tissue engineering, there is a need for quantitative methods to map cell density inside three-dimensional (3-D) bioreactors to assess tissue growth over time. The current cell mapping methods in 2-D cultures are based on optical microscopy. However, optical methods fail in 3-D due to increased opacity of the tissue. We present an approach for measuring the density of cells embedded in a hydrogel to generate quantitative maps of cell density in a living, 3-D tissue culture sample. METHODS:Quantification of cell density was obtained by calibrating the 1H T2, magnetization transfer (MT) and diffusion-weighted nuclear magnetic resonance (NMR) signals to samples of known cell density. Maps of cell density were generated by weighting NMR images by these parameters post-calibration. RESULTS:The highest sensitivity weighting arose from MT experiments, which yielded a limit of detection (LOD) of [Formula: see text] cells/mL/ √{Hz} in a 400 MHz (9.4 T) magnet. CONCLUSION:This mapping technique provides a noninvasive means of visualizing cell growth within optically opaque bioreactors. SIGNIFICANCE:We anticipate that such readouts of tissue culture growth will provide valuable feedback for controlled cell growth in bioreactors.

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