Speckle-Tracking Strain Analysis for Mapping Spatiotemporal Contractility of Induced Pluripotent Stem Cell (iPSC)-Derived Cardiomyocytes.
cardiovascular disease modeling
contractility properties
drug screening
functional assay
human iPSC-derived cardiomyocytes
video microscopy
Journal
Current protocols
ISSN: 2691-1299
Titre abrégé: Curr Protoc
Pays: United States
ID NLM: 101773894
Informations de publication
Date de publication:
Sep 2023
Sep 2023
Historique:
medline:
26
9
2023
pubmed:
25
9
2023
entrez:
25
9
2023
Statut:
ppublish
Résumé
Human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (hiPSC-CMs) hold tremendous potential for cardiovascular disease modeling, drug screening, personalized medicine, and pathophysiology studies. The availability of a robust protocol and functional assay for studying phenotypic behavior of hiPSC-CMs is essential for establishing an in vitro disease model. Many heart diseases manifest due to changes in the mechanical strain of cardiac tissue. Therefore, non-invasive evaluation of the contractility properties of hiPSC-CMs remains crucial to gain an insight into the pathogenesis of cardiac diseases. Speckle tracking-based strain analysis is an efficient non-invasive method that uses video microscopy and image analysis of beating hiPSC-CMs for quantitative evaluation of mechanical contractility properties. This article presents step-by-step protocols for extracting quantitative contractility properties of an hiPSC-CM system obtained from five members of a family, of whom three were affected by DiGeorge syndrome, using speckle tracking-based strain analysis. The hiPSCs from the family members were differentiated and purified into hiPSC-CMs using metabolic selection. Time-lapse images of hiPSC-CMs were acquired using high-spatial-resolution and high-time-resolution phase-contrast video microscopy. Speckled images were characterized by evaluating the cross-correlation coefficient, speckle size, speckle contrast, and speckle quality of the images. The optimum parameters of the speckle tracking algorithm were determined by performing sensitivity analysis concerning computation time, effective mapping area, average contraction velocity, and strain. Furthermore, the hiPSC-CM response to adrenaline was evaluated to validate the sensitivity of the strain analysis algorithm. Then, we applied speckle tracking-based strain analysis to characterize the dynamic behavior of patient-specific hiPSC-CMs from the family members affected/unaffected by DiGeorge syndrome. Here, we report an efficient and manipulation-free method to analyze the contraction displacement vector and velocity field, contraction-relaxation strain rate, and contractile cycles. Implementation of this method allows for quantitative analysis of the contractile phenotype characteristics of hiPSC-CMs to distinguish possible cardiac manifestation of DiGeorge syndrome. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Differentiation of iPSCs into iPSC-derived cardiomyocytes (iPSC-CMs) and metabolic selection of differentiated iPSC-CMs Support Protocol 1: Culture, maintenance, and expansion of human iPSCs Support Protocol 2: Immunohistochemistry of iPSC-CMs Basic Protocol 2: Time-lapse speckle imaging of iPSC-CMs and speckle quality characterization Support Protocol 3: Enhancement of local contrast of videos by applying contrast limited adaptive histogram equalization (CLAHE) to all frames Support Protocol 4: Evaluation of average speckle size Support Protocol 5: Evaluation of average speckle contrast Support Protocol 6: Determination of relative peak height, Pc(x), of consecutive images acquired from video microscopy of iPSC-CMs Basic Protocol 3: Speckle tracking-based analysis of beating iPSC-CMs Support Protocol 7: Validation of sensitivity of the speckle tracking analysis for mapping the contractility of iPSC-CMs Basic Protocol 4: Data extraction, visualization, and mapping of contractile cycles of iPSC-CMs.
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e889Subventions
Organisme : CIHR Foundation Grant
Organisme : Hungarian National Research, Development and Innovation Fund
ID : OTKA K 128369
Organisme : Hungarian National Research, Development and Innovation Fund
ID : GINOP-2.1.1-15-2015-00369
Organisme : K128444, National Cardiovascular Laboratories grant
ID : RRF-2.3.1-21-2022-00003
Organisme : Canadian Research Chair
Organisme : Medicine by Design (University of Toronto)
Informations de copyright
© 2023 Wiley Periodicals LLC.
Références
Azevedo, P. S., Polegato, B. F., Minicucci, M. F., Paiva, S. A., & Zornoff, L. A. (2016). Cardiac remodeling: Concepts, clinical impact, pathophysiological mechanisms and pharmacologic treatment. Arquivos Brasileiros De Cardiologia, 106, 62-69. https://doi.org/10.5935/abc.20160005
Burridge, P. W., Matsa, E., Shukla, P., Lin, Z. C., Churko, J. M., Ebert, A. D., Lan, F., Diecke, S., Huber, B., Mordwinkin, N. M., Plews, J. R., Abilez, O. J., Cui, B., Gold, J. D., & Wu, J. C. (2014). Chemically defined generation of human cardiomyocytes. Nature Methods, 11, 855-860. https://doi.org/10.1038/nmeth.2999
Bellin, M., Marchetto, M. C., Gage, F. H., & Mummery, C. L. (2012). Induced pluripotent stem cells: The new patient? Nature Reviews Molecular Cell Biology, 13, 713-726. https://doi.org/10.1038/nrm3448
Berecz, T., Husvéth-Tóth, M., Mioulane, M., Merkely, B., Apáti, Á., & Földes, G. (2020). Generation and analysis of pluripotent stem cell-derived cardiomyocytes and endothelial cells for high content screening purposes. Methods in Molecular Biology, 2150, 57-77. https://doi.org/10.1007/7651_2019_222
Birket, M. J., Ribeiro, M. C., Kosmidis, G., Ward, D., Leitoguinho, A. R., van de Pol, V., Dambrot, C., Devalla, H. D., Davis, R. P., Mastroberardino, P. G., Atsma, D. E., Passier, R., & Mummery, C. L. (2015). Contractile defect caused by mutation in MYBPC3 revealed under conditions optimized for human PSC-cardiomyocyte function. Cell Reports, 13, 733-745. https://doi.org/10.1016/j.celrep.2015.09.025
Foroosh, H., Zerubia, J. B., & Berthold, M. (2002). Extension of phase correlation to subpixel registration. IEEE Transactions on Image Processing, 11, 188-200. https://doi.org/10.1109/83.988953
Guo, Y., & Pu, W. T. (2020). Cardiomyocyte maturation: New phase in development. Circulation Research, 126, 1086-1106. https://doi.org/10.1161/CIRCRESAHA.119.315862
Izadifar, M., Kelly, M. E., & Peeling, L. (2017). Synchrotron speckle-based x-ray phase-contrast imaging for mapping intra-aneurysmal blood flow without contrast agent. Biomedical Physics & Engineering Express, 4, 015011. https://doi.org/10.1088/2057-1976/aa8e0d
Kelly, D. J., Azeloglu, E. U., Kochupura, P. V., Sharma, G. S., & Gaudette, G. R. (2007). Accuracy and reproducibility of a sub-pixel extended phase correlation method to determine micron level displacements in the heart. Medical Engineering & Physics, 29, 154-162. https://doi.org/10.1016/j.medengphy.2006.01.001
Lemme, M., Ulmer, B. M., Lemoine, M. D., Zech, A. T. L., Flenner, F., Ravens, U., Reichenspurner, H., Rol-Garcia, M., Smith, G., Hansen, A., Christ, T., & Eschenhagen, T. (2018). Atrial-like engineered heart tissue: An in vitro model of the human atrium. Stem Cell Reports, 11, 1378-1390. https://doi.org/10.1016/j.stemcr.2018.10.008
McNally, L. A., Altamimi, T. R., Fulghum, K., & Hill, B. G. (2021). Considerations for using isolated cell systems to understand cardiac metabolism and biology. Journal of Molecular and Cellular Cardiology, 153, 26-41. https://doi.org/10.1016/j.yjmcc.2020.12.007
Park, H., Yeom, E., & Lee, S. J. (2016). X-ray PIV measurement of blood flow in deep vessels of a rat: An in vivo feasibility study. Scientific Reports, 6, 19194. https://doi.org/10.1038/srep19194
Sala, L., van Meer, B. J., Tertoolen, L. G. J., Bakkers, J., Bellin, M., Davis, R. P., Denning, C., Dieben, M. A. E., Eschenhagen, T., Giacomelli, E., Grandela, C., Hansen, A., Holman, E. R., Jongbloed, M. R. M., Kamel, S. M., Koopman, C. D., Lachaud, Q., Mannhardt, I., Mol, M. P. H., … Mummery, C. L. (2018). MUSCLEMOTION: A versatile open software tool to quantify cardiomyocyte and cardiac muscle contraction in vitro and in vivo, Circulation Research, 122, e5-e16. https://doi.org/10.1161/CIRCRESAHA.117.312067
Serrano, R., McKeithan, W. L., Mercola, M., & Carlos del Álamo, J. (2017). High-throughput functional screening assay of force and stiffness in IPSC derived cardiomyocytes. Biophysical Journal, 114, 312A. https://doi.org/10.1016/j.bpj.2017.11.1763
Sleppy, J. (2023). Speckle size via autocorrelation. MATLAB Central File Exchange. Retrieved July 2, 2023, from https://www.mathworks.com/matlabcentral/fileexchange/25046-speckle-size-via-autocorrelation
Takahashi, K., Tanabe, K., Ohnuki, M., Narita, M., Ichisaka, T., Tomoda, K., & Yamanaka, S. (2007). Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell, 131, 861-872. https://doi.org/10.1016/j.cell.2007.11.019
Takahashi, K., & Yamanaka, S. (2006). Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell, 126, 663-676. https://doi.org/10.1016/j.cell.2006.07.024
Yang, X., Rodriguez, M., Pabon, L., Fischer, K. A., Reinecke, H., Regnier, M., Sniadecki, N. J., Ruohola-Baker, H., & Murry, C. E. (2014). Tri-iodo-l-thyronine promotes the maturation of human cardiomyocytes-derived from induced pluripotent stem cells. Journal of Molecular and Cellular Cardiology, 72, 296-304. https://doi.org/10.1016/j.yjmcc.2014.04.005
Zhu, R., Millrod, M. A., Zambidis, E. T., & Tung, L. (2016). Variability of action potentials within and among cardiac cell clusters derived from human embryonic stem cells. Scientific Reports, 6, 18544. https://doi.org/10.1038/srep18544