Low-adhesion culture selection for human iPS cell-derived cardiomyocytes
Tetsutaro Kikuchi1, Katsuhisa Matsuura2, Tatsuya Shimizu2
1Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University (TWIns), 8-1 Kawada-cho, Shinjuku-ku, Tokyo, 162-8666, Japan. kikuchi.tetsutaro@twmu.ac.jp.
Insights
Researchers developed a novel low-adhesion culture method to enrich cardiomyocytes derived from human induced pluripotent stem cells. This technique effectively isolates functional cardiomyocytes, offering a promising alternative to existing selection strategies.
Area of Science:
- Cardiology
- Stem Cell Biology
- Biotechnology
Background:
- Generating cardiomyocytes from human induced pluripotent stem cells (hiPSCs) is crucial for regenerative medicine and disease modeling.
- Current hiPSC-derived cardiomyocyte populations often contain non-contractile cells, requiring efficient selection methods for experimental consistency.
Purpose of the Study:
- To investigate low-adhesion culture as a novel method for enriching cardiomyocytes derived from hiPSCs.
- To compare the efficacy and functional outcomes of low-adhesion culture selection with established cardiomyocyte selection techniques.
Main Methods:
- hiPSC-derived cardiac cells were cultured on low-adhesion surfaces using bovine serum albumin and dextran sulfate.
- Comparison of low-adhesion culture with glucose-free culture and puromycin resistance gene selection.
- Assessment of cardiomyocyte purity, function (pulsation, beta-adrenergic response), and gene expression related to subtype commitment and maturation.
Main Results:
- Low-adhesion culture significantly increased the proportion of cardiac troponin T-positive cardiomyocytes.
- Enriched cardiomyocytes maintained autonomous pulsation and responsiveness to beta-adrenergic stimuli.
- No significant differences in subtype commitment or maturation gene expression were observed compared to other methods.
Conclusions:
- hiPSC-derived cardiomyocytes exhibit greater resistance to low-adhesion culture compared to non-contractile cells.
- Low-adhesion culture selection is a viable and effective alternative for obtaining purified populations of hiPSC-derived cardiomyocytes.
Abstract:
Despite progress in generating cardiomyocytes from pluripotent stem cells, these populations often include non-contractile cells, necessitating cardiomyocyte selection for experimental purpose. This study explores a novel cardiomyocyte enrichment mechanism: low-adhesion culture selection. The cardiac cells derived from human induced pluripotent stem cells were subjected to a coating-free low-adhesion culture using bovine serum albumin and high molecular weight dextran sulfate. This approach effectively increased the population of cardiac troponin T-positive cardiomyocytes. Similar results were obtained with commercially available low-adhesion culture dishes. Subsequently, we accessed the practicality of selection of cardiomyocytes using this phenomenon by comparing it with established methods such as glucose-free culture and selection based on puromycin resistance genes. The cardiomyocytes enriched through low-adhesion culture selection maintained autonomous pulsation and responsiveness to beta-stimuli. Moreover, no significant differences were observed in the expression of genes related to subtype commitment and maturation when compared to other selection methods. In conclusion, cardiomyocytes derived from pluripotent stem cells were more low-adhesion culture resistant than their accompanying non-contractile cells, and low-adhesion culture is an alternative method for selection of pluripotent stem cell-derived cardiomyocytes.
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