Methods in Molecular Biology (2022) 2436: 135–144

DOI 10.1007/7651_2021_432

© Springer Science+Business Media, LLC 2021

Published online: 10 September 2021

In Vitro 3D Mechanical Stimulation to Tendon-Derived Stem

Cells by Bioreactor

Ziming Chen, Peilin Chen, Rui Ruan, and Minghao Zheng

Abstract

Bioreactors can offer an advanced platform to provide conditions that mimic the native microenvironment,

which can also provide stretching environment for mechanobiology research. Tendon-derived stem cells

(TDSCs) are a type of mechanosensitive and multipotent cells, which behave differently in diverse mechan-

ical stretching environments. We have proved the in vitro three-dimensional (3D) mechanical stimulation

could closely mimic the stretching environment in vivo. Thus, here we describe applying a customized

bioreactor to provide 3D force for mechanical stimulation on TDSC in vitro.

Key words Bioreactor, Mechanical loading, Mechanobiology, RT-qPCR, Tendon formation

1

Introduction

Mechanobiology is a multidisciplinary subject including biology,

engineering, and physics, which investigates the influence of

mechanical stress in life science. Cells have the ability to sense

force, and in turn, respond to the extracellular environment

[1]. Thus, mechanobiology is an essential discipline with wide

application prospect.

Various mechanical stimulation types are discussed nowadays,

including compression stimulation, tensile strain, ventilation pres-

sure, flow shear stress, and so forth [2–4]. For tendon, a connective

tissue that attaches muscle to bone, it experiences numerous repet-

itive stretching over a long period of time. Thus, here we use

stretching tensile strain to mimic its nature environment.

Tendon-derived stem cell (TDSC) is a type of mechanosensitive

and multipotent cells, which behaves differently in diverse mechan-

ical stretching environments [5]. Thus, it is an idea source for

mechanobiology study. A proper loading protocol is essential for

tendon

differentiation

of

TDSCs

[6].

Two-dimensional

(2D) loading models, which are applied to monolayer cells, are

the most commonly used for tendon study. However, in our

Ziming Chen and Peilin Chen contributed equally to this work.

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