A versatile pumpless multi-channel fluidics system for maintenance and real-time functional assessment of tissue and cells

Varun Kamat; Matthew K Grumbine; Khang Bao; Kedar Mokate; Gamal Khalil; Daniel Cook; Brandon Clearwater; Richard Hirst; Jarrod Harman; Myriam Boeck; Zhongjie Fu; Lois E H Smith; Moloy Goswami; Thomas J Wubben; Emily M Walker; Jie Zhu; Scott A Soleimanpour; Jarrad M Scarlett; Brian M Robbings; Daniel Hass; James B Hurley; Ian R Sweet

doi:10.1016/j.crmeth.2023.100642

A versatile pumpless multi-channel fluidics system for maintenance and real-time functional assessment of tissue and cells

Cell Rep Methods. 2023 Nov 20;3(11):100642. doi: 10.1016/j.crmeth.2023.100642. Epub 2023 Nov 13.

Authors

Varun Kamat¹, Matthew K Grumbine², Khang Bao², Kedar Mokate¹, Gamal Khalil², Daniel Cook², Brandon Clearwater³, Richard Hirst³, Jarrod Harman⁴, Myriam Boeck⁵, Zhongjie Fu⁴, Lois E H Smith⁴, Moloy Goswami⁶, Thomas J Wubben⁶, Emily M Walker⁷, Jie Zhu⁷, Scott A Soleimanpour⁷, Jarrad M Scarlett⁸, Brian M Robbings⁹, Daniel Hass¹⁰, James B Hurley¹⁰, Ian R Sweet¹¹

Affiliations

¹ University of Washington Medicine Diabetes Institute, University of Washington, Seattle, WA 98109, USA.
² EnTox Sciences, Inc., Mercer Island, WA 98040, USA.
³ Technical Assembly Service Corporation, Seattle, WA 98109, USA.
⁴ Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, Boston, MA 02115, USA.
⁵ Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, Boston, MA 02115, USA; Eye Center, Medical Center, Faculty of Medicine, University of Freiburg, Freiburg im Breisgau, Germany.
⁶ Department of Ophthalmology and Visual Sciences, University of Michigan, Ann Arbor, MI 48105, USA.
⁷ Department of Internal Medicine, University of Michigan, Ann Arbor, MI 98195, USA.
⁸ University of Washington Medicine Diabetes Institute, University of Washington, Seattle, WA 98109, USA; Department of Pediatric Gastroenterology and Hepatology, Seattle Children's Hospital, Seattle, WA 98145, USA.
⁹ University of Washington Medicine Diabetes Institute, University of Washington, Seattle, WA 98109, USA; Department of Biochemistry, University of Washington, Seattle, WA 98109, USA.
¹⁰ Department of Biochemistry, University of Washington, Seattle, WA 98109, USA.
¹¹ University of Washington Medicine Diabetes Institute, University of Washington, Seattle, WA 98109, USA; EnTox Sciences, Inc., Mercer Island, WA 98040, USA. Electronic address: isweet@uw.edu.

Abstract

To address the needs of the life sciences community and the pharmaceutical industry in pre-clinical drug development to both maintain and continuously assess tissue metabolism and function with simple and rapid systems, we improved on the initial BaroFuse to develop it into a fully functional, pumpless, scalable multi-channel fluidics instrument that continuously measures changes in oxygen consumption and other endpoints in response to test compounds. We and several other laboratories assessed it with a wide range of tissue types including retina, pancreatic islets, liver, and hypothalamus with both aqueous and gaseous test compounds. The setup time was less than an hour for all collaborating groups, and there was close agreement between data obtained from the different laboratories. This easy-to-use system reliably generates real-time metabolic and functional data from tissue and cells in response to test compounds that will address a critical need in basic and applied research.

Keywords: BaroFuse; CP: Biotechnology; CP: Metabolism; endothelial cells; hypothalamus; hypoxia; islets; lactate production; liver; oxygen consumption; perifusion; retina.

MeSH terms

Gases / metabolism
Insulin Secretion
Islets of Langerhans* / metabolism
Oxygen / metabolism
Oxygen Consumption

Substances

Oxygen
Gases

Abstract

MeSH terms

Substances

Grants and funding