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

Cell Reports: Methods (Nov 2023)

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

Affiliations

Varun Kamat: University of Washington Medicine Diabetes Institute, University of Washington, Seattle, WA 98109, USA
Matthew K. Grumbine: EnTox Sciences, Inc., Mercer Island, WA 98040, USA
Khang Bao: EnTox Sciences, Inc., Mercer Island, WA 98040, USA
Kedar Mokate: University of Washington Medicine Diabetes Institute, University of Washington, Seattle, WA 98109, USA
Gamal Khalil: EnTox Sciences, Inc., Mercer Island, WA 98040, USA
Daniel Cook: EnTox Sciences, Inc., Mercer Island, WA 98040, USA
Brandon Clearwater: Technical Assembly Service Corporation, Seattle, WA 98109, USA
Richard Hirst: Technical Assembly Service Corporation, Seattle, WA 98109, USA
Jarrod Harman: Department of Ophthalmology, Boston Children’s Hospital, Harvard Medical School, Boston, MA 02115, USA
Myriam Boeck: 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
Zhongjie Fu: Department of Ophthalmology, Boston Children’s Hospital, Harvard Medical School, Boston, MA 02115, USA
Lois E.H. Smith: Department of Ophthalmology, Boston Children’s Hospital, Harvard Medical School, Boston, MA 02115, USA
Moloy Goswami: Department of Ophthalmology and Visual Sciences, University of Michigan, Ann Arbor, MI 48105, USA
Thomas J. Wubben: Department of Ophthalmology and Visual Sciences, University of Michigan, Ann Arbor, MI 48105, USA
Emily M. Walker: Department of Internal Medicine, University of Michigan, Ann Arbor, MI 98195, USA
Jie Zhu: Department of Internal Medicine, University of Michigan, Ann Arbor, MI 98195, USA
Scott A. Soleimanpour: Department of Internal Medicine, University of Michigan, Ann Arbor, MI 98195, USA
Jarrad M. Scarlett: 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
Brian M. Robbings: University of Washington Medicine Diabetes Institute, University of Washington, Seattle, WA 98109, USA; Department of Biochemistry, University of Washington, Seattle, WA 98109, USA
Daniel Hass: Department of Biochemistry, University of Washington, Seattle, WA 98109, USA
James B. Hurley: Department of Biochemistry, University of Washington, Seattle, WA 98109, USA
Ian R. Sweet: University of Washington Medicine Diabetes Institute, University of Washington, Seattle, WA 98109, USA; EnTox Sciences, Inc., Mercer Island, WA 98040, USA; Corresponding author

Journal volume & issue: Vol. 3, no. 11
p. 100642

Abstract

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Summary: 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. Motivation: There is a critical need for developing in vitro methods that provide mechanistic insight at the tissue and intracellular levels as well as to reduce the need for in vivo studies. Flow culture is recognized as superior to static for both maintenance and assessment of tissue, as static systems are unable to maintain viability of most tissue preparations that are not cellular monolayers. However, technical challenges and a need for increased amounts of biological samples are impediments to their routine use in the majority of laboratories in the life sciences. We have addressed these two issues by developing a simple-to-use flow system that utilizes gas pressure instead of peristaltic or syringe pumps to drive multi-channel microflow. The delivery of steady flow over a wide range of flow rates enables small amounts of tissue to be maintained and assessed.

Published in Cell Reports: Methods

ISSN: 2667-2375 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Technology: Chemical technology: Biotechnology; Science: Chemistry: Organic chemistry: Biochemistry
Website: https://www.cell.com/cell-reports-methods

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