Generation of induced pluripotent stem cells and cardiac myocyte-like cells from a patient with noncompaction of ventricular myocardium

Abstract

Objective To obtain induced pluripotent stem cells (iPS) and cardiac myocyte-like cells (CMs) from a patient with noncompaction of ventricular myocardium (NVM). Methods Urine cells from a 16-year-old female patient with NVM and a 24-year-old healthy male volunteer were infected with Sendai virus to obtain iPS cells, which were further induced to differentiate into iPS-CMs. Both the iPS cells and iPS-CMs were identified by immunofluorescence staining, alkaline phosphatase activity, karyotype analysis, embryoid body (EBs) formation and cell patch-clamp experiment. Whole-exome sequencing of the blood sample, iPS cells and iPS-CMs from the NVM patient was performed for genetic comparison to identify gene mutations potentially related with NVM. Results We successfully obtained urine cells, iPS cells and iPS-CMs from the NVM patient and the healthy volunteer. Immunofluorescence assay showed that the iPS cells expressed pluripotent stem cell markers Oct4, Sox2, Ssea4 and TRA-1-81. iPS cells exhibited a high alkaline phosphatase activity with a normal karyotype, and could be induced to differentiate into 3 germ layers in vitro. Immunocytochemical staining revealed that iPS-CMs expressed cardiac myocyte-specific markers TNNT2 and α-actinin. Patch-clamp experiment demonstrated the presence of functional cardiac-specific voltage-gated Na+ currents and action potentials in the iPS-CMs. Whole-exome sequencing showed similar genetic profiles between the blood samples, iPS cells and iPS-CMs of the NVM patient, and several gene mutations were found to have potential associations with NVM. Conclusion We successfully obtain iPS cells and iPS-CMs from the NVM patient, and these cells provide useful cell models for the study of NVM using iPS technology.

Keywords

• noncompaction of ventricular myocardium
• induced pluripotent stem cells
• cardiac myocytes
• urine cells