| Autor: |
Afshar ME; Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Canada.; Donnelly Centre for Cellular and Biomolecular Research, Toronto, Canada., Abraha HY; Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Canada.; Donnelly Centre for Cellular and Biomolecular Research, Toronto, Canada., Bakooshli MA; Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Canada.; Donnelly Centre for Cellular and Biomolecular Research, Toronto, Canada., Davoudi S; Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Canada.; Donnelly Centre for Cellular and Biomolecular Research, Toronto, Canada., Thavandiran N; Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Canada.; Donnelly Centre for Cellular and Biomolecular Research, Toronto, Canada., Tung K; Li Ka Shing Knowledge Institute, St. Michael's Hospital, Toronto, Canada., Ahn H; Li Ka Shing Knowledge Institute, St. Michael's Hospital, Toronto, Canada.; Department of Surgery, University of Toronto, Toronto, Canada., Ginsberg HJ; Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Canada.; Li Ka Shing Knowledge Institute, St. Michael's Hospital, Toronto, Canada.; Department of Surgery, University of Toronto, Toronto, Canada., Zandstra PW; Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Canada.; Donnelly Centre for Cellular and Biomolecular Research, Toronto, Canada.; Michael Smith Laboratories and the School of Biomedical Engineering, University of British Columbia, Vancouver, Canada., Gilbert PM; Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Canada. penney.gilbert@utoronto.ca.; Donnelly Centre for Cellular and Biomolecular Research, Toronto, Canada. penney.gilbert@utoronto.ca.; Department of Biochemistry, University of Toronto, Toronto, Canada. penney.gilbert@utoronto.ca.; Department of Cell and Systems Biology, University of Toronto, Toronto, Canada. penney.gilbert@utoronto.ca. |
| Abstrakt: |
Three-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. |
