A Cylindrical Molding Method for the Biofabrication of Plane-Shaped Skeletal Muscle Tissue.

Minghao Nie,Shoji Takeuchi,Kenta Fukushima,Ai Shima,Yuya Morimoto

doi:10.3390/mi12111411

Abstract

Muscle tissues can be fabricated in vitro by culturing myoblast-populated hydrogels. To counter the shrinkage of the myoblast-populated hydrogels during culture, a pair of anchors are generally utilized to fix the two ends of the hydrogel. Here, we propose an alternative method to counter the shrinkage of the hydrogel and fabricate plane-shaped skeletal muscle tissues. The method forms myoblast-populated hydrogel in a cylindrical cavity with a central pillar, which can prevent tissue shrinkage along the circumferential direction. By eliminating the usages of the anchor pairs, our proposed method can produce plane-shaped skeletal muscle tissues with uniform width and thickness. In experiments, we demonstrate the fabrication of plane-shaped (length: ca. 10 mm, width: 5~15 mm) skeletal muscle tissue with submillimeter thickness. The tissues have uniform shapes and are populated with differentiated muscle cells stained positive for myogenic differentiation markers (i.e., myosin heavy chains). In addition, we show the assembly of subcentimeter-order tissue blocks by stacking the plane-shaped skeletal muscle tissues. The proposed method can be further optimized and scaled up to produce cultured animal products such as cultured meat.

Highlights

The biofabrication of muscle tissue aims at the in vitro reconstruction of muscle tissues, using cells and biomaterials to replicate the compositions, morphologies, and functions of in vivo muscle tissues, which has broad applications in drug development [1,2,3], biohybrid robotics [4,5,6,7,8], regenerative medicine [9,10,11], and cellular agriculture [12,13,14]
The general biofabrication process of muscle tissue starts from the crosslinking of a hydrogel matrix populated with myoblasts, since some types of cells can adhere to the matrix, exerting mechanical forces to and remodeling the matrix [15]
Shape and Homogeneity of the Biofabricated Plane-Shaped Skeletal Muscle Tissues that the length of the fabricated tissue is mainly decided by the peripheral length of the

Summary

Introduction

The biofabrication of muscle tissue aims at the in vitro reconstruction of muscle tissues, using cells and biomaterials to replicate the compositions, morphologies, and functions of in vivo muscle tissues, which has broad applications in drug development [1,2,3], biohybrid robotics [4,5,6,7,8], regenerative medicine [9,10,11], and cellular agriculture [12,13,14]. The general biofabrication process of muscle tissue starts from the crosslinking of a hydrogel matrix populated with myoblasts, since some types of cells (such as fibroblasts and myoblasts) can adhere to the matrix, exerting mechanical forces to and remodeling the matrix [15]. To prevent the over-shrinkage of the myoblast-populated hydrogel matrix, anchoring methods are proposed to reinforce the tissue by fixing the two ends of the myoblast-populated hydrogel using a pair of anchors/pillars [18,19,20]. The anchors can prevent the drastic shrinkage of the myoblast-populated hydrogel but can sustain the tensions generated within the tissue to promote the differentiation of myoblasts and mature the muscle tissues.

Methods

Results

Conclusion