Abstract
BackgroundInduced pluripotent stem cell-derived cardiomyocytes (iPSC-CM) are regarded as promising cell type for cardiac cell replacement therapy, but it is not known whether the developmental stage influences their persistence and functional integration in the host tissue, which are crucial for a long-term therapeutic benefit. To investigate this, we first tested the cell adhesion capability of murine iPSC-CM in vitro at three different time points during the differentiation process and then examined cell persistence and quality of electrical integration in the infarcted myocardium in vivo.MethodsTo test cell adhesion capabilities in vitro, iPSC-CM were seeded on fibronectin-coated cell culture dishes and decellularized ventricular extracellular matrix (ECM) scaffolds. After fixed periods of time, stably attached cells were quantified. For in vivo experiments, murine iPSC-CM expressing enhanced green fluorescent protein was injected into infarcted hearts of adult mice. After 6–7 days, viable ventricular tissue slices were prepared to enable action potential (AP) recordings in transplanted iPSC-CM and surrounding host cardiomyocytes. Afterwards, slices were lysed, and genomic DNA was prepared, which was then used for quantitative real-time PCR to evaluate grafted iPSC-CM count.ResultsThe in vitro results indicated differences in cell adhesion capabilities between day 14, day 16, and day 18 iPSC-CM with day 14 iPSC-CM showing the largest number of attached cells on ECM scaffolds. After intramyocardial injection, day 14 iPSC-CM showed a significant higher cell count compared to day 16 iPSC-CM. AP measurements revealed no significant difference in the quality of electrical integration and only minor differences in AP properties between d14 and d16 iPSC-CM.ConclusionThe results of the present study demonstrate that the developmental stage at the time of transplantation is crucial for the persistence of transplanted iPSC-CM. iPSC-CM at day 14 of differentiation showed the highest persistence after transplantation in vivo, which may be explained by a higher capability to adhere to the extracellular matrix.
Highlights
Induced pluripotent stem cell-derived cardiomyocytes are regarded as promising cell type for cardiac cell replacement therapy, but it is not known whether the developmental stage influences their persistence and functional integration in the host tissue, which are crucial for a long-term therapeutic benefit
The results of the present study demonstrate that the developmental stage at the time of transplantation is crucial for the persistence of transplanted Induced pluripotent stem cell-derived cardiomyocytes (iPSC-CM). Induced pluripotent stem cells (iPSC)-CM at day 14 of differentiation showed the highest persistence after transplantation in vivo, which may be explained by a higher capability to adhere to the extracellular matrix
Cells were transfected with the α-PIG plasmid vector containing the Puromycin N-acetyl-transferase (PAC)-(encoding puromycin N-acetyl-transferase) and Internal ribosomal entry site (IRES) flanked Enhanced green fluorescent protein (eGFP)-gene under control of the α-Myosin Heavy Chain promoter (GenBank Accession No U71441), as described previously [22, 29]. iPS cells were grown on inactivated murine embryonic fibroblasts in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 15% fetal calf serum, 1× non-essential amino acids, 2 mM L-glutamine, 100 μM ß-mercaptoethanol, and 1000 IU/ml leukemia-inhibiting factor (Merck Millipore, Billerica, USA)
Summary
Induced pluripotent stem cell-derived cardiomyocytes (iPSC-CM) are regarded as promising cell type for cardiac cell replacement therapy, but it is not known whether the developmental stage influences their persistence and functional integration in the host tissue, which are crucial for a long-term therapeutic benefit. Experiments in small and large animal models of myocardial infarction (MI) showed safe application and an improved heart function after transplantation of iPSC-CM [8,9,10,11,12]. As result of these preclinical studies, the first clinical trials using human iPSC-CM were recently approved by regulatory authorities [13]. The mechanisms behind cell homing into recipient cardiac tissue or cell loss are poorly understood and the poor cell persistence after transplantation remains an unsolved challenge
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
