Microfluidic Cardiac Cell Culture Model (μCCCM)

Abstract

Physiological heart development and cardiac function rely on the response of cardiac cells to mechanical stress during hemodynamic loading and unloading. These stresses, especially if sustained, can induce changes in cell structure, contractile function, and gene expression. Current cell culture techniques commonly fail to adequately replicate physical loading observed in the native heart. Therefore, there is a need for physiologically relevant in vitro models that recreate mechanical loading conditions seen in both normal and pathological conditions. To fulfill this need, we have developed a microfluidic cardiac cell culture model (μCCCM) that for the first time allows in vitro hemodynamic stimulation of cardiomyocytes by directly coupling cell structure and function with fluid induced loading. Cells are cultured in a small (1 cm diameter) cell culture chamber on a thin flexible silicone membrane. Integrating the cell culture chamber with a pump, collapsible pulsatile valve and an adjustable resistance element (hemostatic valve) in series allow replication of various loading conditions experienced in the heart. This paper details the design, modeling, fabrication and characterization of fluid flow, pressure and stretch generated at various frequencies to mimic hemodynamic conditions associated with the normal and failing heart. Proof-of-concept studies demonstrate successful culture of an embryonic cardiomyoblast line (H9c2 cells) and establishment of an in vivo like phenotype within this system.

An integrated microfluidic array system for evaluating toxicity and teratogenicity of drugs on embryonic zebrafish developmental dynamics

“…Recently, Giridharan et al reported that the fluid shear stress was significant ͑10 dynes/ cm 2 ͒ at the height of 200 m when the well was 250 m in depth. 47 However, the exposed shear stress, at the same height in the well of 2 mm in depth, was about 0, regardless of whether the fluid shear stress on the order was more than 10 dynes/ cm 2 or not. In this work, the positioned spherical embryo was attached on the bottom surface of culture chamber with a height approximately 500 m, whereas the well was as high as 1700 m. Additionally, the flow rate ͑24 mm/s͒ presented in the above mentioned model was much higher than that ͑4 l / min͒ used in our tests.…”

Microfluidic Organ‐on‐a‐Chip Technology for Advancement of Drug Development and Toxicology

“…From there, the fl ow would travel in the blood, reaching the kidney, heart, bone marrow, brain, spleen, and vasculature. Reproduced with permission: lung, [ 26 ] Copyright 2011, Springer; gut, [ 40 ] Copyright 2012, The Royal Society of Chemistry; liver, [ 16 ] Copyright 2013, Elsevier; kidney, [ 50 ] Copyright 2013, The Royal Society of Chemistry; heart, [ 58 ] Copyright 2010, American Chemical Society; bone marrow, [ 67 ] Copyright 2014, Nature Publishing Group; brain, [ 69 ] Copyright 2012, Springer; spleen, [ 70 ] Copyright 2014, The Royal Society of Chemistry; and vasculature, [ 66 ] Copyright 2011, American Institute of Physics. Characterizing the toxicity of several compounds using liver co-culture and SMART-scale analysis HepG2/C3A and MDCK [20] Drug metabolism using liver and intestinal slices Harvested rat liver and intestinal slices [22] Testing the effects of ethanol-induced toxicity on liver slices Harvested rat liver slices [21] Functional analysis Creating model with bile canaliculi formation Harvested rat hepatocytes [14] Incorporated biosensors for pH monitoring in chip HepG2 [15] The kinetic reactions representing metabolism, measured and compared with mathematical model…”

“…[ 56 ] Due to the advantages of 3D microfl uidic systems, there has been a shift towards using these designs to represent cardiac situations. [ 57 ] Giridharan et al [ 58 ] developed a model designed to replicate physical loading in the left ventricle. This microfl uidic cardiac cell culture model (µCCCM) contains a cell culture chamber, a pump, a collapsible pulsatile valve, and an adjustable hemostatic valve.…”

Effects of hydraulic pressure on cardiomyoblasts in a microfluidic device

“…Recently, Giridharan et al developed a microfluidic cardiac cell culture model (lCCCM) that incorporated pulsatile stretch and pressure on cardiac cells in a single device. 11 The mechanical stimulations can mimic various physiological conditions, including heart failure, hypertension, and hypotension. However, the structure and manipulation of this model device is too complicated for daily operations in a common biological laboratory.…”