GENERATION, CULTIVATION AND CHARACTERIZATION OF LARGE SCALE STEM CELL DERIVED BIOARTIFICIAL CARDIAC TISSUE

Generation of a large-scale stem cell derived bioartificial cardiac tissue (BCT) as a potential therapeutic option to replace damaged myocardium after myocardial infarction. Large-scale BCT was generated from cardiomyocytes (CMs) derived from human induced pluripotent stem cells (hiPSC) mixed with h...

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Bibliographic Details
Published in:International journal of artificial organs 2019-08, Vol.42 (8)
Main Authors: Elena, R S M, Szepes, M, Melchert, A, Halloin, C, Zweigerdt, R, Gruh, I
Format: Article
Language:English
Subjects:
Bioreactors
Calcium
Cardiomyocytes
Cultivation
Fibroblasts
Genetic code
Heart
Hydrogels
Immunofluorescence
Integration
Maturation
Myocardial infarction
Myocardium
Pluripotency
Proteins
Rhythms
Staining
Stem cells
Striations
Tissues
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Summary:Generation of a large-scale stem cell derived bioartificial cardiac tissue (BCT) as a potential therapeutic option to replace damaged myocardium after myocardial infarction. Large-scale BCT was generated from cardiomyocytes (CMs) derived from human induced pluripotent stem cells (hiPSC) mixed with human fibroblasts and a hydrogel solution in a newly designed tissue chamber with a diameter of 43 mm. The BCT development was microscopically recorded and evaluated. Maturation of CMs within the tissue was confirmed by immunofluorescence staining for cardiac markers (α-SA, cTnT) as well as cell junction proteins (Con. 43, N-Cad). The integration of the tissue chamber in the novel custom-made bioreactor allowed investigating the initial parameters for electromechanical stimulation (such as pressure, stress amplitude, frequency) necessary for proper tissue maturation in a more physiological manner. To investigate the calcium handling through the tissue a new transgenic hiPSC line containing a genetically encoded calcium indicator (GCaMP6f) has been developed. Microscopically evaluation at different time points reveals a progressive reduction of nearly ~27% of the initial volume after 15 days, ccorresponding with an increased intercellular connectivity. After 48h, the tissue exhibited spontaneous contractions and coordinated and rhythmic contraction within the whole tissue after 6 days. As preliminary analysis, the beating of the tissue was video-optically recorded showing an estimated beating rate of 102 beats per minute. Immunofluorescence staining performed at day 15 showed highly organised cross-striations of sarcomere proteins. Initial integration of the tissue chamber in the bioreactor and implementation of the new GCaMP6f reporter cell line is still on-going. The initial results confirmed that is possible to generate a large-scale BCT. However, the properties of the tissue have to be further investigated.
ISSN:0391-3988
1724-6040