Micro Vacuum Chuck and Tensile Test System for Bio-Mechanical Evaluation of 3D Tissue Constructed of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes (hiPS-CM).

Kaoru Uesugi,Keisuke Morishima,Fumiaki Shima,Ken Fukumoto,Shigeru Miyagawa,Mitsuru Akashi,Yoshinari Tsukamoto,Takami Akagi,Ayami Hiura,Yoshiki Sawa

doi:10.3390/mi10070487

Kaoru Uesugi, Keisuke Morishima + Show 8 more

Open Access

https://doi.org/10.3390/mi10070487

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Journal: Micromachines	Publication Date: Jul 19, 2019
Citations: 11	License type: CC BY 4.0

Affiliation: Osaka University

Abstract

In this report, we propose a micro vacuum chuck (MVC) which can connect three-dimensional (3D) tissues to a tensile test system by vacuum pressure. Because the MVC fixes the 3D tissue by vacuum pressure generated on multiple vacuum holes, it is expected that the MVC can fix 3D tissue to the system easily and mitigate the damage which can happen by handling during fixing. In order to decide optimum conditions for the size of the vacuum holes and the vacuum pressure, various sized vacuum holes and vacuum pressures were applied to a normal human cardiac fibroblast 3D tissue. From the results, we confirmed that a square shape with 100 µm sides was better for fixing the 3D tissue. Then we mounted our developed MVCs on a specially developed tensile test system and measured the bio-mechanical property (beating force) of cardiac 3D tissue which was constructed of human induced pluripotent stem cell-derived cardiomyocytes (hiPS-CM); the 3D tissue had been assembled by the layer-by-layer (LbL) method. We measured the beating force of the cardiac 3D tissue and confirmed the measured force followed the Frank-Starling relationship. This indicates that the beating property of cardiac 3D tissue obtained by the LbL method was close to that of native cardiac tissue.

Highlights

IntroductionThree-dimensional (3D) tissues which are constructed by cells in the environment in vitro have been applied in a wide range of fields such as regenerative medicine [1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25,26], drug development [2,9,11,12,13,14,15,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41,42], disease modelling for pathology [11,12,13,14,15,36,43,44,45,46], bioactuators [47,48,49,50,51,52], food industry [42,52,53,54,55,56], and BioArt [52,55,56,57,58,59,60,61,62]
We developed a micro vacuum chuck (MVC) which could fix the 3D tissue by vacuum pressure and we developed a special tensile test system for it
In order to optimize the size of the vacuum holes and vacuum pressure, we applied various sized vacuum holes and vacuum pressures to a NHCF 3D tissue

Summary

Introduction

Three-dimensional (3D) tissues which are constructed by cells in the environment in vitro have been applied in a wide range of fields such as regenerative medicine [1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25,26], drug development [2,9,11,12,13,14,15,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41,42], disease modelling for pathology [11,12,13,14,15,36,43,44,45,46], bioactuators [47,48,49,50,51,52], food industry [42,52,53,54,55,56], and BioArt [52,55,56,57,58,59,60,61,62]. With the advancement of 3D tissue technologies, evaluation methods for them have been demanded Analytical approaches such as biochemical, immunological, morphological [24,63], electrophysiological, and motion image analysis [31] methods have been applied to evaluate artificial tissues. These methods, cannot evaluate bio-mechanical properties directly. In the field of regenerative medicine, bio-mechanical evaluations are important for constructing 3D tissues which replicate living tissues. Because the sense of touch mainly depends on mechanical information, the mechanical properties of artificial 3D tissues must be known

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