title: Culture and Sampling of Primary Adipose Tissue in Practical Microfluidic Systems.
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.
authors.affiliation: Department of Chemistry and Biochemistry, Auburn University, 179 Chemistry Bldg, Auburn, AL, 36849, USA.
authors.forename: Jessica C
authors.initials: JC
authors.lastname: Brooks
authors.affiliation: Department of Anatomy Physiology and Pharmacology, Auburn University, Auburn, AL, 36849, USA.
authors.forename: Robert L
authors.initials: RL
authors.lastname: Judd
authors.affiliation: Department of Chemistry and Biochemistry, Auburn University, 179 Chemistry Bldg, Auburn, AL, 36849, USA. chris.easley@auburn.edu.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Easley
mesh_terms: D017667:Adipocytes / Q000166:cytology / Q000378:metabolism; D000273:Adipose Tissue / Q000166:cytology / Q000378:metabolism; D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D004867:Equipment Design; D056656:Lab-On-A-Chip Devices; D008297:Male; D051379:Mice; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D044085:Microfluidics / Q000295:instrumentation*
keywords: 3D printing; Adipose tissue; Cell culture; Explants; Free fatty acids; Glycerol; Hormones; Insulin signaling; Microfluidics; Secretion