Effective derivation of ventricular cardiomyocytes from hPSCs using ascorbic acid-containing maturation medium

Area of Science:

• Stem cell biology
• Cardiovascular research
• Regenerative medicine

Background:

• Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) are valuable for disease modeling, drug screening, and cardiac repair.
• Efficient maturation and subtype specification of hPSC-CMs are crucial for their clinical and research applications.

Purpose of the Study:

• To develop an optimized method for selecting and maturing hPSC-CMs into specific subtypes.
• To investigate the role of glucose starvation, nutrition complex, and ascorbic acid in hPSC-CM maturation.
• To compare gene expression patterns under different maturation conditions using next-generation sequencing (NGS).

Main Methods:

• hPSC differentiation guided by Wnt signaling regulation.
• Optimization of selection and maturation medium under glucose starvation conditions.
• Supplementation with either a nutrition complex or ascorbic acid.
• Quantification of cardiac Troponin T (cTnT)-positive cells.
• Next-generation sequencing (NGS) analysis of cardiomyocyte-specific gene expression.

Main Results:

• Albumin and ascorbic acid supplementation yielded more cTnT-positive cardiomyocytes compared to B27.
• Ascorbic acid enriched the maturation of ventricular cardiomyocytes.
• NGS analysis revealed distinct gene expression patterns under varied selection and maturation conditions.

Conclusions:

• The optimized selection and maturation protocol enhances hPSC-CM maturation and subtype specification.
• This method offers a simple and efficient approach for generating specific cardiomyocyte subtypes.
• The findings facilitate advancements in biomedical research and the development of cell-based cardiac therapies.