Label-Free Raman Spectroscopy for Assessing Purity and Maturation of hiPSC-Derived Cardiac Tissue

Area of Science:

• Biotechnology
• Stem Cell Biology
• Cardiovascular Research

Background:

• Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are vital for drug toxicity evaluation, discovery, and heart failure treatment.
• Large-scale hiPSC-CM production necessitates robust methods for quality, purity, and maturation assessment.
• A demand exists for label-free, non-damaging techniques to characterize cell molecular profiles.

Purpose of the Study:

• To employ label-free Raman microscopy for evaluating hiPSC-derived CMs.
• To investigate the correlation between specific molecular components (cytochrome c, myoglobin) and CM purity/maturation.
• To validate Raman spectroscopy as a quality control tool for hiPSC-CMs.

Main Methods:

• Utilized label-free Raman microscopy to characterize hiPSC-derived CMs.
• Investigated Raman spectroscopy of cytochrome c and myoglobin for correlation with purity and maturation.
• Validated findings using mixtures of cardiomyocytes and fibroblasts, and hiPSC-CMs of varying efficiencies.
• Compared Raman profiles of immature and matured CMs after rapid pacing.

Main Results:

• Raman spectroscopy of cytochrome c and myoglobin correlated with hiPSC-CM purity and maturation.
• Validation experiments confirmed the correlation in mixed cell populations and varying hiPSC-CM efficiencies.
• Matured hiPSC-CMs displayed distinct Raman profiles compared to immature ones.

Conclusions:

• Raman spectroscopy of specific components accurately reflects hiPSC-CM purity and maturation.
• Label-free Raman microscopy is a promising nondestructive, high-content, and efficient method for hiPSC-CM quality control.
• This technique can enhance the reliability of hiPSC-CMs in drug discovery and regenerative medicine.