A Versatile Pumpless Multi-channel Fluidics System for Maintenance and Real-time Functional Assessment of Tissue and Cells

Overview

Journal Cell Rep Methods

Specialty Cell Biology

Date 2023 Nov 14

PMID 37963464

Authors

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

Soon will be listed here.

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.

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References

Sweet I, Gilbert M, Scott S, Todorov I, Jensen R, Nair I . Glucose-stimulated increment in oxygen consumption rate as a standardized test of human islet quality. Am J Transplant. 2007; 8(1):183-92. DOI: 10.1111/j.1600-6143.2007.02041.x. View

Doliba N, Qin W, Vatamaniuk M, Buettger C, Collins H, Magnuson M . Cholinergic regulation of fuel-induced hormone secretion and respiration of SUR1-/- mouse islets. Am J Physiol Endocrinol Metab. 2006; 291(3):E525-35. DOI: 10.1152/ajpendo.00579.2005. View

Chan G, Kleinheinz T, Peterson D, Moffat J . A simple high-content cell cycle assay reveals frequent discrepancies between cell number and ATP and MTS proliferation assays. PLoS One. 2013; 8(5):e63583. PMC: 3656927. DOI: 10.1371/journal.pone.0063583. View

Grumbine M, Kamat V, Bao K, Crupi T, Mokate K, Lim R . Maintaining and Assessing Various Tissue and Cell Types of the Eye Using a Novel Pumpless Fluidics System. J Vis Exp. 2023; (197). PMC: 10791547. DOI: 10.3791/65399. View

Paish H, Reed L, Brown H, Bryan M, Govaere O, Leslie J . A Bioreactor Technology for Modeling Fibrosis in Human and Rodent Precision-Cut Liver Slices. Hepatology. 2019; 70(4):1377-1391. PMC: 6852483. DOI: 10.1002/hep.30651. View

Nolfi-Donegan D, Braganza A, Shiva S . Mitochondrial electron transport chain: Oxidative phosphorylation, oxidant production, and methods of measurement. Redox Biol. 2020; 37:101674. PMC: 7767752. DOI: 10.1016/j.redox.2020.101674. View

Brandhorst H, Brandhorst D, Hering B, Bretzel R . Significant progress in porcine islet mass isolation utilizing liberase HI for enzymatic low-temperature pancreas digestion. Transplantation. 1999; 68(3):355-61. DOI: 10.1097/00007890-199908150-00006. View

Quent V, Loessner D, Friis T, Reichert J, Hutmacher D . Discrepancies between metabolic activity and DNA content as tool to assess cell proliferation in cancer research. J Cell Mol Med. 2010; 14(4):1003-13. PMC: 3823131. DOI: 10.1111/j.1582-4934.2010.01013.x. View

Cook D, Hales C . Intracellular ATP directly blocks K+ channels in pancreatic B-cells. Nature. 1984; 311(5983):271-3. DOI: 10.1038/311271a0. View

10.

Rountree A, Karkamkar A, Khalil G, Folch A, Cook D, Sweet I . BaroFuse, a novel pressure-driven, adjustable-throughput perfusion system for tissue maintenance and assessment. Heliyon. 2016; 2(12):e00210. PMC: 5155043. DOI: 10.1016/j.heliyon.2016.e00210. View

11.

Ono J, Lacy P, Michael H, Greider M . Studies of the functional and morphologic status of islets maintained at 24 C for four weeks in vitro. Am J Pathol. 1979; 97(3):489-503. PMC: 2042423. View

12.

Soleimanpour S, Gupta A, Bakay M, Ferrari A, Groff D, Fadista J . The diabetes susceptibility gene Clec16a regulates mitophagy. Cell. 2014; 157(7):1577-90. PMC: 4184276. DOI: 10.1016/j.cell.2014.05.016. View

13.

Schulze T, Morsi M, Reckers K, Bruning D, Seemann N, Panten U . Metabolic amplification of insulin secretion is differentially desensitized by depolarization in the absence of exogenous fuels. Metabolism. 2017; 67:1-13. DOI: 10.1016/j.metabol.2016.10.008. View

14.

Leung D, Chu S . Measurement of Oxidative Stress: Mitochondrial Function Using the Seahorse System. Methods Mol Biol. 2017; 1710:285-293. DOI: 10.1007/978-1-4939-7498-6_22. View

15.

Bisbach C, Hass D, Robbings B, Rountree A, Sadilek M, Sweet I . Succinate Can Shuttle Reducing Power from the Hypoxic Retina to the O-Rich Pigment Epithelium. Cell Rep. 2020; 31(5):107606. PMC: 7273505. DOI: 10.1016/j.celrep.2020.107606. View

16.

Sweet I, Cook D, Wiseman R, Greenbaum C, Lernmark A, Matsumoto S . Dynamic perifusion to maintain and assess isolated pancreatic islets. Diabetes Technol Ther. 2002; 4(1):67-76. DOI: 10.1089/15209150252924111. View

17.

Masand S, Mignone L, Zahn J, Shreiber D . Nanoporous membrane-sealed microfluidic devices for improved cell viability. Biomed Microdevices. 2011; 13(6):955-61. DOI: 10.1007/s10544-011-9565-z. View

18.

Goncalves I, Carvalho V, Rodrigues R, Pinho D, Teixeira S, Moita A . Organ-on-a-Chip Platforms for Drug Screening and Delivery in Tumor Cells: A Systematic Review. Cancers (Basel). 2022; 14(4). PMC: 8870045. DOI: 10.3390/cancers14040935. View

19.

Du J, Rountree A, Cleghorn W, Contreras L, Lindsay K, Sadilek M . Phototransduction Influences Metabolic Flux and Nucleotide Metabolism in Mouse Retina. J Biol Chem. 2015; 291(9):4698-710. PMC: 4813492. DOI: 10.1074/jbc.M115.698985. View

20.

Hirtenstein M, Clark J, Lindgren G, Vretblad P . Microcarriers for animal cell culture: a brief review of theory and practice. Dev Biol Stand. 1980; 46:109-16. View