Microfluidics-enabled 96-well Perfusion System for High-throughput Tissue Engineering and Long-term All-optical Electrophysiology

Overview

Journal Lab Chip

Publisher Royal Society of Chemistry

Specialties Biotechnology
Chemistry

Date 2020 Sep 30

PMID 32996969

Citations 16

Authors

Lai Wei

Weizhen Li

Emilia Entcheva

Zhenyu Li

Affiliations

Soon will be listed here.

Abstract

This work demonstrates a novel high-throughput (HT) microfluidics-enabled uninterrupted perfusion system (HT-μUPS) and validates its use with chronic all-optical electrophysiology in human excitable cells. HT-μUPS consists of a soft multichannel microfluidic plate cover which could button on a commercial HT 96-well plate. Herein, we demonstrate the manufacturing process of the system and its usages in acute and chronic all-optical electrophysiological studies of human induced pluripotent stem-cell-derived cardiomyocytes (iPSC-CM) and engineered excitable (spiking HEK) cells. HT-μUPS perfusion maintained functional voltage and calcium responses in iPSC-CM and spiking HEK cells under spontaneous conditions and under optogenetic pacing. Long-term culture with HT-μUPS improved cell viability and optogenetically-tracked calcium responses in spiking HEK cells. The simplicity of this design and its compatibility with HT all-optical electrophysiology can empower cell-based assays for personalized medicine using patient-derived cells.

Citing Articles

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References

Kuzmiak-Glancy S, Covian R, Femnou A, Glancy B, Jaimes 3rd R, Wengrowski A . Cardiac performance is limited by oxygen delivery to the mitochondria in the crystalloid-perfused working heart. Am J Physiol Heart Circ Physiol. 2017; 314(4):H704-H715. PMC: 5966767. DOI: 10.1152/ajpheart.00321.2017. View

Karbassi E, Fenix A, Marchiano S, Muraoka N, Nakamura K, Yang X . Cardiomyocyte maturation: advances in knowledge and implications for regenerative medicine. Nat Rev Cardiol. 2020; 17(6):341-359. PMC: 7239749. DOI: 10.1038/s41569-019-0331-x. View

Nagel G, Szellas T, Huhn W, Kateriya S, Adeishvili N, Berthold P . Channelrhodopsin-2, a directly light-gated cation-selective membrane channel. Proc Natl Acad Sci U S A. 2003; 100(24):13940-5. PMC: 283525. DOI: 10.1073/pnas.1936192100. View

Magdy T, Schuldt A, Wu J, Bernstein D, Burridge P . Human Induced Pluripotent Stem Cell (hiPSC)-Derived Cells to Assess Drug Cardiotoxicity: Opportunities and Problems. Annu Rev Pharmacol Toxicol. 2017; 58:83-103. PMC: 7386286. DOI: 10.1146/annurev-pharmtox-010617-053110. View

Huang Y, Walker A, Miller E . A Photostable Silicon Rhodamine Platform for Optical Voltage Sensing. J Am Chem Soc. 2015; 137(33):10767-76. PMC: 4666802. DOI: 10.1021/jacs.5b06644. View

Klimas A, Ortiz G, Boggess S, Miller E, Entcheva E . Multimodal on-axis platform for all-optical electrophysiology with near-infrared probes in human stem-cell-derived cardiomyocytes. Prog Biophys Mol Biol. 2019; 154:62-70. PMC: 6728218. DOI: 10.1016/j.pbiomolbio.2019.02.004. View

Jackman C, Carlson A, Bursac N . Dynamic culture yields engineered myocardium with near-adult functional output. Biomaterials. 2016; 111:66-79. PMC: 5074846. DOI: 10.1016/j.biomaterials.2016.09.024. View

Bhatia S, Ingber D . Microfluidic organs-on-chips. Nat Biotechnol. 2014; 32(8):760-72. DOI: 10.1038/nbt.2989. View

Yoshimitsu R, Hattori K, Sugiura S, Kondo Y, Yamada R, Tachikawa S . Microfluidic perfusion culture of human induced pluripotent stem cells under fully defined culture conditions. Biotechnol Bioeng. 2013; 111(5):937-47. DOI: 10.1002/bit.25150. View

10.

Ruan J, Tulloch N, Razumova M, Saiget M, Muskheli V, Pabon L . Mechanical Stress Conditioning and Electrical Stimulation Promote Contractility and Force Maturation of Induced Pluripotent Stem Cell-Derived Human Cardiac Tissue. Circulation. 2016; 134(20):1557-1567. PMC: 5123912. DOI: 10.1161/CIRCULATIONAHA.114.014998. View

11.

Entcheva E, Bub G . All-optical control of cardiac excitation: combined high-resolution optogenetic actuation and optical mapping. J Physiol. 2016; 594(9):2503-10. PMC: 4850200. DOI: 10.1113/JP271559. View

12.

Park J, Werley C, Venkatachalam V, Kralj J, Dib-Hajj S, Waxman S . Screening fluorescent voltage indicators with spontaneously spiking HEK cells. PLoS One. 2014; 8(12):e85221. PMC: 3877367. DOI: 10.1371/journal.pone.0085221. View

13.

Rolland J, van Dam R, Schorzman D, Quake S, DeSimone J . Solvent-resistant photocurable liquid fluoropolymers for microfluidic device fabrication [corrected]. J Am Chem Soc. 2004; 126(8):2322-3. DOI: 10.1021/ja031657y. View

14.

Pierce C, Uppuluri P, Tristan A, Wormley Jr F, Mowat E, Ramage G . A simple and reproducible 96-well plate-based method for the formation of fungal biofilms and its application to antifungal susceptibility testing. Nat Protoc. 2008; 3(9):1494-500. PMC: 2741160. DOI: 10.1038/nport.2008.141. View

15.

Park S, Mondal K, Treadway 3rd R, Kumar V, Ma S, Holbery J . Silicones for Stretchable and Durable Soft Devices: Beyond Sylgard-184. ACS Appl Mater Interfaces. 2018; 10(13):11261-11268. DOI: 10.1021/acsami.7b18394. View

16.

Glauche F, John G, Arain S, Knepper A, Neubauer A, Goelling D . Toward Microbioreactor Arrays: A Slow-Responding Oxygen Sensor for Monitoring of Microbial Cultures in Standard 96-Well Plates. J Lab Autom. 2015; 20(4):438-46. DOI: 10.1177/2211068215573924. View

17.

Zhou J, Ellis A, Voelcker N . Recent developments in PDMS surface modification for microfluidic devices. Electrophoresis. 2009; 31(1):2-16. DOI: 10.1002/elps.200900475. View

18.

Hirt M, Boeddinghaus J, Mitchell A, Schaaf S, Bornchen C, Muller C . Functional improvement and maturation of rat and human engineered heart tissue by chronic electrical stimulation. J Mol Cell Cardiol. 2014; 74:151-61. DOI: 10.1016/j.yjmcc.2014.05.009. View

19.

Chen S, Hung P, Lee P . Microfluidic array for three-dimensional perfusion culture of human mammary epithelial cells. Biomed Microdevices. 2011; 13(4):753-8. DOI: 10.1007/s10544-011-9545-3. View

20.

Unger M, Chou H, Thorsen T, Scherer A, Quake S . Monolithic microfabricated valves and pumps by multilayer soft lithography. Science. 2001; 288(5463):113-6. DOI: 10.1126/science.288.5463.113. View