A Micromanipulation‐Actuated Large‐Scale Screening to Identify Optimized Microphysiological Model Parameters in Skeletal Muscle Regeneration

Abstract Hydrogel‐based 3D cell cultures are extensively utilized to create biomimetic cellular microstructures. However, there is still lack of effective method for both evaluation of the complex interaction of cells with hydrogel and the functionality of the resulting micro‐structures. This limitation impedes the further application of these microstructures as microphysiological models (microPMs) for the screening of potential culture condition combinations to enhance the skeletal muscle regeneration. This paper introduces a two‐probe micromanipulation method for the large‐scale assessment of viscoelasticity and contractile force (CF) of skeletal muscle microPMs, which are produced in high‐throughput via microfluidic spinning and 96‐well culture. The collected data demonstrate that viscoelasticity parameters (E^* and tanδ) and CF both measured in a solution environment are indicative of the formation of cellular structures without hydrogel residue and the subsequent generation of myotubes, respectively. This study have developed screening criterias that integrate E^*, tanδ, and CF to examine the effects of multifactorial interactions on muscle fiber repair under hypoxic conditions and within bioprinted bipennate muscle structures. This approach has improved the quality of hypoxic threshold evaluation and aligned cell growth in 3D. The proposed method is useful in exploring the role of different factors in muscle tissue regeneration with limited resources.