Lactate- and immunomagnetic-purified hiPSC-derived cardiomyocytes generate comparable engineered cardiac tissue...

Area of Science:

• Stem cell biology
• Cardiovascular research
• Proteomics

Background:

• Three-dimensional engineered cardiac tissue (ECT) from human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) is a valuable model for cardiac studies.
• Previous research suggested metabolic (lactate) purification of hiPSC-CMs induces an ischemic cardiomyopathy-like phenotype, contrasting with magnetic antibody-based cell sorting (MACS) purification.

Purpose of the Study:

• To investigate whether lactate purification versus MACS purification of hiPSC-CMs impacts the properties of resultant hiPSC-ECTs.
• To determine if lactate purification leads to irreversible phenotypic changes in hiPSC-CMs used for ECT generation.

Main Methods:

• hiPSC-CMs were differentiated and purified using either lactate-based media or MACS.
• Purified hiPSC-CMs were integrated into 3D hiPSC-ECTs and cultured for 4 weeks.
• Structural (sarcomere length), functional (isometric twitch force, Ca2+ transients), and proteomic analyses (global and quantitative proteomics) were performed.

Main Results:

• Global proteomics revealed differential phenotypes between lactate- and MACS-purified hiPSC-CMs prior to ECT formation.
• No significant differences were observed in sarcomere length, isometric twitch force, or Ca2+ transient measurements between lactate- and MACS-derived hiPSC-ECTs.
• Quantitative proteomics showed no significant differences in protein pathway expression or myofilament proteoforms in the final hiPSC-ECTs.

Conclusions:

• hiPSC-CMs purified via lactate or MACS yield ECTs with comparable structural, functional, and proteomic characteristics.
• Lactate purification does not appear to induce an irreversible detrimental phenotype in hiPSC-CMs used for generating functional engineered cardiac tissue.