Dispersible oxygen microsensors map oxygen gradients in three-dimensional cell cultures

Sasha Cai Lesher-Pérez; Ge-Ah Kim; Chuan-hsien Kuo; Brendan M. Leung; Sanda Mong; Taisuke Kojima; Christopher Moraes; M. D. Thouless; Gary D. Luker; Shuichi Takayama

doi:10.1039/C7BM00119C

Dispersible oxygen microsensors map oxygen gradients in three-dimensional cell cultures†

Sasha Cai Lesher-Pérez,

^abk Ge-Ah Kim,^bc Chuan-hsien Kuo,^a Brendan M. Leung,^ab Sanda Mong,^a Taisuke Kojima,^bd Christopher Moraes,

^abe M. D. Thouless,^cf Gary D. Luker^aghi and Shuichi Takayama

*^abdj

Author affiliations

* Corresponding authors

^a Department of Biomedical Engineering, University of Michigan, Ann Arbor, USA
E-mail: takayama@umich.edu

^b Biointerfaces Institute, University of Michigan, Ann Arbor, USA

^c Department of Materials Science and Engineering, University of Michigan, Ann Arbor, USA

^d Department of Macromolecular Science and Engineering, University of Michigan, Ann Arbor, USA

^e Department of Chemical Engineering, McGill University, Montreal, Canada

^f Department of Mechanical Engineering, University of Michigan, Ann Arbor, USA

^g Department of Radiology, University of Michigan School of Medicine, Ann Arbor, MI, USA

^h Center for Molecular Imaging, Department of Radiology, University of Michigan, Ann Arbor, MI, USA

ⁱ Department of Microbiology and Immunology, University of Michigan, Ann Arbor, USA

^j Michigan Center for Integrative Research in Critical Care, University of Michigan, Ann Arbor, USA

^k Chemical and Biomolecular Engineering Department, University of California, Los Angeles, USA

Abstract

Phase fluorimetry, unlike the more commonly used intensity-based measurement, is not affected by differences in light paths from culture vessels or by optical attenuation through dense 3D cell cultures and hydrogels thereby minimizing dependence on signal intensity for accurate measurements. This work describes the use of phase fluorimetry on oxygen-sensor microbeads to perform oxygen measurements in different microtissue culture environments. In one example, cell spheroids were observed to deplete oxygen from the cell-culture medium filling the bottom of conventional microwells within minutes, whereas oxygen concentrations remained close to ambient levels for several days in hanging-drop cultures. By dispersing multiple oxygen microsensors in cell-laden hydrogels, we also mapped cell-generated oxygen gradients. The spatial oxygen mapping was sufficiently precise to enable the use of computational models of oxygen diffusion and uptake to give estimates of the cellular oxygen uptake rate and the half-saturation constant. The results show the importance of integrated design and analysis of 3D cell cultures from both biomaterial and oxygen supply aspects. While this paper specifically tests spheroids and cell-laden gel cultures, the described methods should be useful for measuring pericellular oxygen concentrations in a variety of biomaterials and culture formats.

Biomaterials Science

Dispersible oxygen microsensors map oxygen gradients in three-dimensional cell cultures†

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Dispersible oxygen microsensors map oxygen gradients in three-dimensional cell cultures

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