Toward a Microencapsulated 3D hiPSC-Derived in vitro Cardiac Microtissue for Recapitulation of Human Heart Microenvironment Features

Abecasis, Bernardo; Canhão, Pedro G M; Almeida, Henrique V.; Calmeiro, Tomás; Fortunato, Elvira; Gomes‐Alves, Patrícia; Serra, Margarida; Alves, Paula M.

doi:10.3389/fbioe.2020.580744

Cited by 11 publications

(12 citation statements)

References 40 publications

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“…The aggregate metabolic activity was quantified, as previously described by our group, 34 with PrestoBlue (Life Technologies), following the manufacturer's instructions.…”

Section: Methodsmentioning

confidence: 99%

“…The samples were cryosectioned and the immunofluorescence performed as previously described by our group. 34 The primary antibodies used were cardiac troponin T (cTnT) (Thermo Fisher Scientific, diluted 1:200), sarcomeric α-actinin (Sigma-Aldrich, diluted 1:200), collagen I (abcam, diluted 1:100), α-smooth muscle actin (αSMA; Agilent Technologies/DAKO, diluted 1:100), and vimentin (abcam, diluted 1:100). The secondary antibodies used were goat antirabbit IgG Alexa Fluor 486 and goat antimouse Alexa Fluor 594 (all from Thermo Fisher Scientific, diluted 1:500).…”

Section: Methodsmentioning

confidence: 99%

“…The hiPSC-CM aggregates were collected at day 28 and dissociated with TrypLE (37 °C for 5 min) for flow cytometry analysis as previously described by our group. 34 The following conjugated antibodies were used: SIRPα/β (CD172a/b-PE, BioLegend, diluted 1:20 in DPBS) and isotype control IgG2, k-PE (BD Biosciences, diluted 1:5 in DPBS). Stained cell suspensions were analyzed in a BD FACSCalibur cytometer (BD Biosciences), and the analysis was performed with FlowJo software (TreeStar).…”

Section: Methodsmentioning

confidence: 99%

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Human Extracellular-Matrix Functionalization of 3D hiPSC-Based Cardiac Tissues Improves Cardiomyocyte Maturation

Almeida

Tenreiro

Louro

et al. 2021

ACS Appl. Bio Mater.

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Human induced pluripotent stem cells (hiPSC) possess significant therapeutic potential due to their high self-renewal capability and potential to differentiate into specialized cells such as cardiomyocytes. However, generated hiPSCderived cardiomyocytes (hiPSC-CM) are still immature, with phenotypic and functional features resembling the fetal rather than their adult counterparts, which limits their application in cell-based therapies, in vitro cardiac disease modeling, and drug cardiotoxicity screening. Recent discoveries have demonstrated the potential of the extracellular matrix (ECM) as a critical regulator in development, homeostasis, and injury of the cardiac microenvironment. Within this context, this work aimed to assess the impact of human cardiac ECM in the phenotype and maturation features of hiPSC-CM. Human ECM was isolated from myocardium tissue through a physical decellularization approach. The cardiac tissue decellularization process reduced DNA content significantly while maintaining ECM composition in terms of sulfated glycosaminoglycans (s-GAG) and collagen content. These ECM particles were successfully incorporated in three-dimensional (3D) hiPSC-CM aggregates (CM+ECM) with no impact on viability and metabolic activity throughout 20 days in 3D culture conditions. Also, CM+ECM aggregates displayed organized and longer sarcomeres, with improved calcium handling when compared to hiPSC-CM aggregates. This study shows that human cardiac ECM functionalization of hiPSC-based cardiac tissues improves cardiomyocyte maturation. The knowledge generated herein provides essential insights to streamline the application of ECM in the development of hiPSC-based therapies targeting cardiac diseases.

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“…The aggregate metabolic activity was quantified, as previously described by our group, 34 with PrestoBlue (Life Technologies), following the manufacturer's instructions.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Human Extracellular-Matrix Functionalization of 3D hiPSC-Based Cardiac Tissues Improves Cardiomyocyte Maturation

Almeida

Tenreiro

Louro

et al. 2021

ACS Appl. Bio Mater.

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“…[ 76 ] The same applies to other polysaccharide‐based hydrogels such as alginate and hyaluronic acid . [ 189 , 190 , 191 ]…”

Section: Multiparametric Materials Functionalitymentioning

confidence: 99%

Multiparametric Material Functionality of Microtissue‐Based In Vitro Models as Alternatives to Animal Testing

2022

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With the definition of the 3R principle by Russel and Burch in 1959, the search for an adequate substitute for animal testing has become one of the most important tasks and challenges of this time, not only from an ethical, but also from a scientific, economic, and legal point of view. Microtissue-based in vitro model systems offer a valuable approach to address this issue by accounting for the complexity of natural tissues in a simplified manner. To increase the functionality of these model systems and thus make their use as a substitute for animal testing more likely in the future, the fundamentals need to be continuously improved. Corresponding requirements exist in the development of multifunctional, hydrogel-based materials, whose properties are considered in this review under the aspects of processability, adaptivity, biocompatibility, and stability/degradability.

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“…Moreover, the resulting cardiomyocytes exhibited a well-developed and organized sarcomeric structure that collectively indicated improved cardiac maturation. When microencapsulated cardiomyocytes differentiated from iPSC were co-cultured with endothelial cells and stromal cells in Gly-Arg-Gly-Asp-Ser-Pro (GRGDSP) peptide-coupled alginate hydrogel, they achieved structural maturation after 15 days of culture, as evidenced by the presence of matured myofibril alignment accompanied by elongated and well-organized sarcomeres, which were absent in the control cardiomyocytes aggregates ( Abecasis et al, 2020 ). The resulting model also demonstrated dose-dependent response towards known cardiotoxins such as doxorubicin.…”

Section: Moving Towards Clinical Translationmentioning

confidence: 99%

Polysaccharide-Based Hydrogels for Microencapsulation of Stem Cells in Regenerative Medicine

Lee

Khoo

et al. 2021

Front. Bioeng. Biotechnol.

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Stem cell-based therapy appears as a promising strategy to induce regeneration of damaged and diseased tissues. However, low survival, poor engraftment and a lack of site-specificity are major drawbacks. Polysaccharide hydrogels can address these issues and offer several advantages as cell delivery vehicles. They have become very popular due to their unique properties such as high-water content, biocompatibility, biodegradability and flexibility. Polysaccharide polymers can be physically or chemically crosslinked to construct biomimetic hydrogels. Their resemblance to living tissues mimics the native three-dimensional extracellular matrix and supports stem cell survival, proliferation and differentiation. Given the intricate nature of communication between hydrogels and stem cells, understanding their interaction is crucial. Cells are incorporated with polysaccharide hydrogels using various microencapsulation techniques, allowing generation of more relevant models and further enhancement of stem cell therapies. This paper provides a comprehensive review of human stem cells and polysaccharide hydrogels most used in regenerative medicine. The recent and advanced stem cell microencapsulation techniques, which include extrusion, emulsion, lithography, microfluidics, superhydrophobic surfaces and bioprinting, are described. This review also discusses current progress in clinical translation of stem-cell encapsulated polysaccharide hydrogels for cell delivery and disease modeling (drug testing and discovery) with focuses on musculoskeletal, nervous, cardiac and cancerous tissues.

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Toward a Microencapsulated 3D hiPSC-Derived in vitro Cardiac Microtissue for Recapitulation of Human Heart Microenvironment Features

Cited by 11 publications

References 40 publications

Human Extracellular-Matrix Functionalization of 3D hiPSC-Based Cardiac Tissues Improves Cardiomyocyte Maturation

Human Extracellular-Matrix Functionalization of 3D hiPSC-Based Cardiac Tissues Improves Cardiomyocyte Maturation

Multiparametric Material Functionality of Microtissue‐Based In Vitro Models as Alternatives to Animal Testing

Polysaccharide-Based Hydrogels for Microencapsulation of Stem Cells in Regenerative Medicine

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