Skip to Content
Merck
  • A systemic evaluation of cardiac differentiation from mRNA reprogrammed human induced pluripotent stem cells.

A systemic evaluation of cardiac differentiation from mRNA reprogrammed human induced pluripotent stem cells.

PloS one (2014-07-30)
Ashish Mehta, Vinod Verma, Manasi Nandihalli, Chrishan J A Ramachandra, Glen L Sequiera, Yuliansa Sudibyo, Yingying Chung, William Sun, Winston Shim
ABSTRACT

Genetically unmodified cardiomyocytes mandated for cardiac regenerative therapy is conceivable by "foot-print free" reprogramming of somatic cells to induced pluripotent stem cells (iPSC). In this study, we report generation of foot-print free hiPSC through messenger RNA (mRNA) based reprograming. Subsequently, we characterize cardiomyocytes derived from these hiPSC using molecular and electrophysiological methods to characterize their applicability for regenerative medicine. Our results demonstrate that mRNA-iPSCs differentiate ontogenetically into cardiomyocytes with increased expression of early commitment markers of mesoderm, cardiac mesoderm, followed by cardiac specific transcriptional and sarcomeric structural and ion channel genes. Furthermore, these cardiomyocytes stained positively for sarcomeric and ion channel proteins. Based on multi-electrode array (MEA) recordings, these mRNA-hiPSC derived cardiomyocytes responded predictably to various pharmacologically active drugs that target adrenergic, sodium, calcium and potassium channels. The cardiomyocytes responded chronotropically to isoproterenol in a dose dependent manner, inotropic activity of nifidipine decreased spontaneous contractions. Moreover, Sotalol and E-4031 prolonged QT intervals, while TTX reduced sodium influx. Our results for the first time show a systemic evaluation based on molecular, structural and functional properties of cardiomyocytes differentiated from mRNA-iPSC. These results, coupled with feasibility of generating patient-specific iPSCs hold great promise for the development of large-scale generation of clinical grade cardiomyocytes for cardiac regenerative medicine.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Ethidium bromide solution, 10 mg/mL
Pricing and availability is not currently available.
Supelco
Nifedipine, Pharmaceutical Secondary Standard; Certified Reference Material
Pricing and availability is not currently available.
Sigma-Aldrich
Ethidium bromide solution, for fluorescence, ~1% in H2O
Pricing and availability is not currently available.
Sigma-Aldrich
Nifedipine, ≥98% (HPLC), powder
Pricing and availability is not currently available.
USP
Nifedipine, United States Pharmacopeia (USP) Reference Standard
Pricing and availability is not currently available.
Sigma-Aldrich
Ethidium bromide solution, BioReagent, for molecular biology, 10 mg/mL in H2O
Pricing and availability is not currently available.
Sigma-Aldrich
Ethidium bromide solution, BioReagent, for molecular biology, 500 μg/mL in H2O
Pricing and availability is not currently available.
Nifedipine, European Pharmacopoeia (EP) Reference Standard
Pricing and availability is not currently available.
Sigma-Aldrich
Ethidium bromide, ≥95.0%
Pricing and availability is not currently available.
Sigma-Aldrich
Ethidium bromide, BioReagent, for molecular biology, powder
Pricing and availability is not currently available.
Sigma-Aldrich
Ethidium bromide, ~95% (HPLC)
Pricing and availability is not currently available.