Speaker
Description
Introduction
Cardiovascular disease (CVD) is the most mortality disease globally, and the effective method of its treatment is still unknown. For this reason, there is growing interest in the use of human induced pluripotent stem cells (iPSCs), which can differentiate into all cells in the human body, including induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) which have the potential for future use in regenerative medicine and cardiology [1]. Additionally, it has no appropriate cellular models' mimetic cardiac tissue; therefore, new methods of studying cardiac cell function are still needed. A technique that can potentially produce cellular models that mimic cardiac tissue function is tissue engineering (TE). TE is designed to create a suitable scaffold to mimic the extracellular matrix (ECM). Currently, numerous studies are conducted using nanofibers as a scaffold for culture cardiac cells. However, there is still a lack of research evaluating the impact of polymer nanofibers on the functions of co-culture cardiac cells with stem cells [2].
Methods
We produced polyurethane (PU) nanofibrous mats (Figure 1A) by using the solution blow spinning (SBS) method. Nanofibrous mats were modified with oxygen plasma and protein solutions (0.01% Matrigel solution) to improve cell adhesion. Here, we studied how nanomaterials influence the function of damaged human cardiomyocytes in co-culture with stem cells. First, cell viability (calcein-AM assay) was checked for 3 day-culture (Figure 1B). Next, immunostaining of the levels of OCT4 protein, which regulates the work of induced pluripotent stem cells, and cTnT2 protein involved in the regulation of cardiomyocytes were conducted after 10 days (Figure 2A and B). To confirm the changes occurring in the co-cultures indicative of the differentiation of iPSCs into cardiomyocytes, gene expression analysis of GATA4 (encoding GATA4), TNNI3 (encoding troponin I), SERCA2 (encoding calcium ATPase-type P-ATPase) were performed after 24h and 10 days (Figure 2C).
Results and Discussion
The study checked the viability of the cells, which confirmed that the co-cultures had a high viability (Figure 2). The changes at the protein and gene level (Figure 2) demonstrate that for cultures on polystyrene plates, iPSC cells can differentiate into cardiomyocytes both under iPSC-CMs in normoxia and when iPSC-CMs were subjected to hypoxia. However, at this stage, further studies are required to confirm whether nanofibrous mats can be used for 3D cellular models to study the regeneration of damaged iPSC-CMs. In addition, studies were conducted on immature iPSC-CMs, and further prospects would be to see how mature cardiomyocytes function after hypoxia and in co-culture with iPSCs on nanofibrous mats.
References
[1] J. Gorecka et al., ‘The potential and limitations of induced pluripotent stem cells to achieve wound healing,’ Stem Cell Res Ther, 2019.
[2] S.N.H. Karimi, “Tri-layered alginate/poly(ε-caprolactone) electrospun scaffold for cardiac tissue engineering,” Polymer International, 2022
Acknowledgment
This work was realized with the frame of project SONATA BIS 2019/34 / E / ST5 / 00381.
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