A Systemized Approach to Investigate Ca(2+) Synchronization in Clusters of Human Induced Pluripotent Stem-Cell...

Area of Science:

• Cardiovascular Research
• Stem Cell Biology
• Regenerative Medicine

Background:

• Induced pluripotent stem cell-derived cardiomyocytes (IPS-CM) hold promise for cardiac repair.
• Current IPS-CM production methods exhibit variability and low reproducibility.
• Understanding factors influencing IPS-CM maturation is critical.

Purpose of the Study:

• To investigate the impact of embryoid body (EB) vs. "on plate" culture on IPS-CM maturation.
• To analyze spontaneous activity and calcium (Ca2+) synchronization in IPS-CM clusters.
• To determine how early maturation stages affect IPS-CM phenotype and response.

Main Methods:

• Systematic investigation of IPS-CM maturation at different time points.
• Comparison of calcium (Ca2+) dynamics in EB-derived vs. "on plate" cultured IPS-CM.
• Analysis of spontaneous activity and regional Ca2+ synchronization.

Main Results:

• Disaggregation of EBs between 0.5-2 weeks yielded spontaneously beating IPS-CM with high Ca2+ synchronization.
• IPS-CM from older EBs (>2 weeks) lacked spontaneous activity and synchronization.
• "On plate" culture of active IPS-CM clusters led to reduced synchronization and loss of spontaneous beating.
• Elevated extracellular Ca2+ revealed distinct Ca2+ handling behaviors in quiescent IPS-CM, traceable to derivation methods.

Conclusions:

• Early maturation interventions, specifically EB disaggregation timing, can predictably influence IPS-CM phenotype.
• Maturation route significantly impacts IPS-CM spontaneous activity and Ca2+ synchronization.
• These findings offer a pathway to enhance the reliability and efficacy of IPS-CM for cardiac regeneration.