Journal Article

ISL1 Protein Transduction Promotes Cardiomyocyte Differentiation from Human Embryonic Stem Cells

Fattahi, Faranak, Fonoudi, Hananeh, Yeganeh, Meghdad, Mojarad, Bahareh Adhami, Ghazizadeh, Zaniar, Rassouli, Hassan, Salekdeh, Ghasem Hosseini, Baharvand, Hossein, Alikhani, Mehdi, Aghdami, Nasser

2013

Overview

Human embryonic stem cells (hESCs) have the potential to provide an unlimited source of cardiomyocytes, which are invaluable resources for drug or toxicology screening, medical research, and cell therapy. Currently a number of obstacles exist such as the insufficient efficiency of differentiation protocols, which should be overcome before hESC-derived cardiomyocytes can be used for clinical applications. Although the differentiation efficiency can be improved by the genetic manipulation of hESCs to over-express cardiac-specific transcription factors, these differentiated cells are not safe enough to be applied in cell therapy. Protein transduction has been demonstrated as an alternative approach for increasing the efficiency of hESCs differentiation toward cardiomyocytes. We present an efficient protocol for the differentiation of hESCs in suspension by direct introduction of a LIM homeodomain transcription factor, Islet1 (ISL1) recombinant protein into the cells. We found that the highest beating clusters were derived by continuous treatment of hESCs with 40 µg/ml recombinant ISL1 protein during days 1-8 after the initiation of differentiation. The treatment resulted in up to a 3-fold increase in the number of beating areas. In addition, the number of cells that expressed cardiac specific markers (cTnT, CONNEXIN 43, ACTININ, and GATA4) doubled. This protocol was also reproducible for another hESC line. This study has presented a new, efficient, and reproducible procedure for cardiomyocytes differentiation. Our results will pave the way for scaled up and controlled differentiation of hESCs to be used for biomedical applications in a bioreactor culture system.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Actinin / Agricultural biotechnology / Biocompatibility / Biology / Biomedical materials / Bioreactors / Cardiomyocytes / Cell culture / Cell Differentiation - drug effects / Cell Differentiation - genetics / Cell Line / Connexin 43 / Developmental biology / Differentiation / DNA binding proteins / Education / Efficiency / Embryo cells / Embryonic stem cells / Embryonic Stem Cells - cytology / Embryonic Stem Cells - drug effects / Embryonic Stem Cells - metabolism / Gene Expression / Genetic engineering / Health screening / Heart / Heart diseases / Homeobox / Humans / Islet-1 protein / LIM-Homeodomain Proteins - genetics / LIM-Homeodomain Proteins - metabolism / LIM-Homeodomain Proteins - pharmacology / Medical research / Myocytes, Cardiac - cytology / Myocytes, Cardiac - drug effects / Myocytes, Cardiac - metabolism / Permeability / Protein Stability / Proteins / Recombinant Fusion Proteins / Recombinant proteins / Science / Security management / Stem cells / Therapeutic applications / Therapy / Toxicology / Transcription (Genetics) / Transcription factors / Transcription Factors - genetics / Transcription Factors - metabolism / Transcription Factors - pharmacology