Abstract
Microfluidic culture of primary adipose tissue allows for reduced sample and reagent volumes as well as constant media perfusion of the cells. By continuously flowing media over the tissue, microfluidic sampling systems can more accurately mimic vascular flow in vivo. Quantitative measurements can be performed on or off chip to provide time-resolved secretion data, furthering insight into the dynamics of the function of adipose tissue. Buoyancy resulting from the large lipid storage capacity in this tissue presents a unique challenge for culture, and it is important to account for this buoyancy during microdevice design. Herein, we describe approaches for microfluidic device fabrication that utilize 3D-printed interface templating to help counteract cell buoyancy. We apply such methods to the culture of both isolated, dispersed primary adipocytes and epididymal adipose explants. To facilitate more widespread adoption of the methodology, the devices presented here are designed for user-friendly operation. Only handheld syringes are needed to control flow, and devices are inexpensive and disposable.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Lee JN, Park C, Whitesides GM (2003) Solvent compatibility of poly(dimethylsiloxane)-based microfluidic devices. Anal Chem 75:6544–6554
Whitesides GM, Ostuni E, Takayama S et al (2001) Soft lithography in biology and biochemistry. Annu Rev Biomed Eng 3:335–373
Easley CJ, Karlinsey JM, Landers JP (2006) On-chip pressure injection for integration of infrared-mediated DNA amplification with electrophoretic separation. Lab Chip 6:601–610
Kim J, Johnson M, Hill P, Gale BK (2009) Microfluidic sample preparation: cell lysis and nucleic acid purification. Integr Biol 1:574–586
Gao Y, Majumdar D, Jovanovic B et al (2011) A versatile valve-enabled microfluidic cell co-culture platform and demonstration of its applications to neurobiology and cancer biology. Biomed Microdevices 13:539–548
Halldorsson S, Lucumi E, Gómez-Sjöberg R, Fleming RMT (2015) Advantages and challenges of microfluidic cell culture in polydimethylsiloxane devices. Biosens Bioelectron 63:218–231
Bhatia SN, Ingber DE (2014) Microfluidic organs-on-chips. Nat Biotechnol 32:760–772
Dishinger JF, Reid KR, Kennedy RT (2009) Quantitative monitoring of insulin secretion from single islets of Langerhans in parallel on a microfluidic chip. Anal Chem 81:3119–3127
Adewola AF, Wang Y, Harvat T et al (2010) A multi-parametric islet perifusion system within a microfluidic perifusion device. J Vis Exp 35:e1649
Yi L, Wang X, Dhumpa R et al (2015) Integrated perfusion and separation systems for entrainment of insulin secretion from islets of Langerhans. Lab Chip 15:823–832
Easley CJ, Rocheleau JV, Head WS, Piston DW (2009) Quantitative measurement of zinc secretion from pancreatic islets with high temporal resolution using droplet-based microfluidics. Anal Chem 81:9086–9095
Godwin LA, Pilkerton ME, Deal KS et al (2011) Passively operated microfluidic device for stimulation and secretion sampling of single pancreatic islets. Anal Chem 83:7166–7172
Brooks JC, Ford KI, Holder DH et al (2016) Macro-to-micro interfacing to microfluidic channels using 3D-printed templates: Application to time-resolved secretion sampling of endocrine tissue. Analyst 141:5714–5721
Moraes C, Labuz JM, Leung BM et al (2013) On being the right size: scaling effects in designing a human-on-a-chip. Integr Biol 5:1149–1161
Godwin LA, Brooks JC, Hoepfner LD et al (2015) A microfluidic interface for the culture and sampling of adiponectin from primary adipocytes. Analyst 140:1019–1025
Erickstad M, Gutierrez E, Groisman A (2015) A low-cost low-maintenance ultraviolet lithography light source based on light-emitting diodes. Lab Chip 15:57–61. doi:10.1039/c4lc00472h
Acknowledgments
Support for the work was provided by the National Institutes of Health (R01 DK093810) as well as by the Department of Chemistry and Biochemistry and the College of Science and Mathematics at Auburn University. The authors would like to thank Mark D. Holtan and Tesfagebriel Hagos for assistance with device fabrication photographs. We also extend thanks to Dr. Leah A. Godwin for initiating the work in this area.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Brooks, J.C., Judd, R.L., Easley, C.J. (2017). Culture and Sampling of Primary Adipose Tissue in Practical Microfluidic Systems. In: Wu, J. (eds) Thermogenic Fat. Methods in Molecular Biology, vol 1566. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6820-6_18
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6820-6_18
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6819-0
Online ISBN: 978-1-4939-6820-6
eBook Packages: Springer Protocols