Influence of Diameter and Cyclic Mechanical Stimulation on the Beating Frequency of Myocardial Cell-Laden Fibers
Abstract
:1. Introduction
2. Results and Discussion
2.1. Cell Infiltration and Proliferation
2.2. iPSC-CM Markers for the BioPacer
2.3. Beating Frequency
2.4. Glucose and Lactate Measurements
2.5. Discussion
3. Conclusions
4. Methods
4.1. Cylindrical Fibrin Gel Constructs
4.2. Cell Culturing
4.3. Bioreactor Set-Up
4.4. Glucose and Lactate Measurements
4.5. Immunohistochemistry
4.6. Beating Frequency
4.7. Quantification of the Markers Content
4.8. Statistics
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Sample Type | Fiber Number | [iPSC-CMs] (Cells/Fiber) | [HUVSMCs] (Cells/Fiber) | [HUVECs] (Cells/Fiber) |
---|---|---|---|---|
Low cell density 500 µm (G1) | 2 | 150,000 | 60,000 | 20,000 |
Mix 500 µm (G2) | 1 | 300,000 | 60,000 | 20,000 |
500 µm (G3) | 2 | 300,000 | 60,000 | 20,000 |
500 µm stretch (G4) | 2 | 300,000 | 60,000 | 20,000 |
2 × 500 µm (G5) | 2 | 300,000 | ||
3 × 500 µm (G6) | 3 | 300,000 | ||
1 mm (G7) | 2 | 500,000 | 120,000 | 60,000 |
1 mm stretch (G8) | 2 | 500,000 | 120,000 | 60,000 |
Mix 1 mm (G9) | 1 | 500,000 | 120,000 | 60,000 |
Antibody | Dilution | Supply |
---|---|---|
Anti-α-Actinin (sarcomeric), mouse monoclonal | 1:50 | Sigma-Aldrich |
Connexin 43 (polyclonal) | 1:50 | Thermofischer |
Connexin 43 Monoclonal Antibody (3D8A5) | 1:50 | Thermofischer |
Anti-CD31 (PECAM-1), mouse monoclonal | 1:100 | Sigma-Aldrich |
CD31 rabbit polyclonal | 1:100 | Abbiotec |
α-smooth muscle actin (monoclonal, clone 1A4) | 1:1000 | Sigma-Aldrich |
Anti-Collagen I antibody (ab34710) | 1:200 | Abcam |
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Kyriakou, S.; Lubig, A.; Sandhoff, C.A.; Kuhn, Y.; Jockenhoevel, S. Influence of Diameter and Cyclic Mechanical Stimulation on the Beating Frequency of Myocardial Cell-Laden Fibers. Gels 2023, 9, 677. https://doi.org/10.3390/gels9090677
Kyriakou S, Lubig A, Sandhoff CA, Kuhn Y, Jockenhoevel S. Influence of Diameter and Cyclic Mechanical Stimulation on the Beating Frequency of Myocardial Cell-Laden Fibers. Gels. 2023; 9(9):677. https://doi.org/10.3390/gels9090677
Chicago/Turabian StyleKyriakou, Stavroula, Andreas Lubig, Cilia A. Sandhoff, Yasmin Kuhn, and Stefan Jockenhoevel. 2023. "Influence of Diameter and Cyclic Mechanical Stimulation on the Beating Frequency of Myocardial Cell-Laden Fibers" Gels 9, no. 9: 677. https://doi.org/10.3390/gels9090677