A 96-well culture platform enables longitudinal analyses of engineered human skeletal muscle microtissue strength

Author:

Afshar Mohammad E.,Abraha Haben Y.,Bakooshli Mohsen A.,Davoudi Sadegh,Thavandiran Nimalan,Tung Kayee,Ahn Henry,Ginsberg Howard J.,Zandstra Peter W.,Gilbert Penney M.

Abstract

AbstractThree-dimensional (3D) in vitro models of human skeletal muscle mimic aspects of native tissue structure and function, thereby providing a promising system for disease modeling, drug discovery or pre-clinical validation, and toxicity testing. Widespread adoption of this research approach is hindered by the lack of easy-to-use platforms that are simple to fabricate and that yield arrays of human skeletal muscle micro-tissues (hMMTs) in culture with reproducible physiological responses that can be assayed non-invasively. Here, we describe a design and methods to generate a reusable mold to fabricate a 96-well platform, referred to as MyoTACTIC, that enables bulk production of 3D hMMTs. All 96-wells and all well features are cast in a single step from the reusable mold. Non-invasive calcium transient and contractile force measurements are performed on hMMTs directly in MyoTACTIC, and unbiased force analysis occurs by a custom automated algorithm, allowing for longitudinal studies of function. Characterizations of MyoTACTIC and resulting hMMTs confirms the capability of the device to support formation of hMMTs that recapitulate biological responses. We show that hMMT contractile force mirrors expected responses to compounds shown by others to decrease (dexamethasone, cerivastatin) or increase (IGF-1) skeletal muscle strength. Since MyoTACTIC supports hMMT long-term culture, we evaluated direct influences of pancreatic cancer chemotherapeutics agents on contraction competent human skeletal muscle myotubes. A single application of a clinically relevant dose of Irinotecan decreased hMMT contractile force generation, while clear effects on myotube atrophy were observed histologically only at a higher dose. This suggests an off-target effect that may contribute to cancer associated muscle wasting, and highlights the value of the MyoTACTIC platform to non-invasively predict modulators of human skeletal muscle function.

Funder

Gouvernement du Canada | Natural Sciences and Engineering Research Council of Canada

Ontario Graduate Scholarship

Ontario Graduate Scholarship Toronto Musculoskeletal Centre Fellowship

Heart and Stroke Foundation of Canada

Gouvernement du Canada | Canadian Institutes of Health Research

CIHR

Canada First Research Excellence Fund

Canada Research Chair Program

Ontario Institute for Regenerative Medicine

Canada Research Chair Program Ontario Research Fund Krembil Foundation

Publisher

Springer Science and Business Media LLC

Subject

Multidisciplinary

Reference75 articles.

1. Periasamy, M. et al. Role of SERCA Pump in Muscle Thermogenesis and Metabolism. Compr. Physiol. 7, 879–890 (2017).

2. Lauretani, F. et al. Age-associated changes in skeletal muscles and their effect on mobility: an operational diagnosis of sarcopenia. J. Appl. Physiol. Bethesda Md 1985 95, 1851–1860 (2003).

3. Maltzahn, J., von, Renaud, J.-M., Parise, G. & Rudnicki, M. A. Wnt7a treatment ameliorates muscular dystrophy. Proc. Natl. Acad. Sci. 109, 20614–20619 (2012).

4. Thomason, D. B. & Booth, F. W. Atrophy of the soleus muscle by hindlimb unweighting. J. Appl. Physiol. Bethesda Md 1985 68, 1–12 (1990).

5. Gaschen, F. P. et al. Dystrophin deficiency causes lethal muscle hypertrophy in cats. J. Neurol. Sci. 110, 149–159 (1992).

同舟云学术

1.学者识别学者识别

2.学术分析学术分析

3.人才评估人才评估

"同舟云学术"是以全球学者为主线,采集、加工和组织学术论文而形成的新型学术文献查询和分析系统,可以对全球学者进行文献检索和人才价值评估。用户可以通过关注某些学科领域的顶尖人物而持续追踪该领域的学科进展和研究前沿。经过近期的数据扩容,当前同舟云学术共收录了国内外主流学术期刊6万余种,收集的期刊论文及会议论文总量共计约1.5亿篇,并以每天添加12000余篇中外论文的速度递增。我们也可以为用户提供个性化、定制化的学者数据。欢迎来电咨询!咨询电话:010-8811{复制后删除}0370

www.globalauthorid.com

TOP

Copyright © 2019-2024 北京同舟云网络信息技术有限公司
京公网安备11010802033243号  京ICP备18003416号-3