Key result
NKX2.5 GFP reporter human iPSCs successfully differentiate into functional cardiac fibroblasts matching patient-derived cells.
Why the study?
In human direct cardiac reprogramming studies, the lack of reporter fibroblasts has hindered screening of factors and development of robust reprogramming protocols.
The creation of a human NKX2.5GFP reporter hiPSC line provides an unlimited cellular model to facilitate research and screening of factors for direct cardiac reprogramming.
Offers scalable iPSC model for fibroblast research; leaves open translation to clinical reprogramming therapies.
Direct cardiac reprogramming has emerged as an interesting approach for the treatment and regeneration of damaged hearts through the direct conversion of fibroblasts into cardiomyocytes or cardiovascular progenitors. However, in studies with human cells, the lack of reporter fibroblasts has hindered the screening of factors and consequently, the development of robust direct cardiac reprogramming protocols.In this study, we have generated functional human NKX2.5 GFP reporter cardiac fibroblasts. We first established a new NKX2.5 GFP reporter human induced pluripotent stem cell (hiPSC) line using a CRISPR-Cas9-based knock-in approach in order to preserve function which could alter the biology of the cells. The reporter was found to faithfully track NKX2.5 expressing cells in differentiated NKX2.5 GFP hiPSC and the potential of NKX2.5-GFP + cells to give rise to the expected cardiac lineages, including functional ventricular- and atrial-like cardiomyocytes, was demonstrated. Then NKX2.5 GFP cardiac fibroblasts were obtained through directed differentiation, and these showed typical fibroblast-like morphology, a specific marker expression profile and, more importantly, functionality similar to patient-derived cardiac fibroblasts. The advantage of using this approach is that it offers an unlimited supply of cellular models for research in cardiac reprogramming, and since NKX2.5 is expressed not only in cardiomyocytes but also in cardiovascular precursors, the detection of both induced cell types would be possible. These reporter lines will be useful tools for human direct cardiac reprogramming research and progress in this field.
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López-Muneta et al. (2022) studied functional cardiac fibroblasts. NKX2.5 GFP reporter human iPSCs was evaluated on Differentiation into functional cardiac fibroblasts. NKX2.5 GFP reporter human iPSCs successfully differentiated into functional cardiac fibroblasts demonstrating typical morphology and expression profiles similar to patient-derived cardiac fibroblasts.
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