MiR-499 Responsive Lethal Construct for Removal of Human Embryonic Stem Cells after Cardiac Differentiation

Deriving cell populations from human embryonic stem cells (hESCs) for cell-based therapy is considered a promising strategy to achieve functional cells, yet its translation to clinical practice depends on achieving fully defined differentiated cells. In this work, we generated a miRNA-responsive let...

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Bibliographic Details
Published in:Scientific reports 2019-10, Vol.9 (1), p.14490-14, Article 14490
Main Authors: Elovic, Edan, Etzion, Sharon, Cohen, Smadar
Format: Article
Language:English
Subjects:
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Animals
Apoptosis
Apoptosis - genetics
bcl-2-Associated X Protein - genetics
Calcium
Cardiomyocytes
Cell culture
Cell differentiation
Cell Differentiation - genetics
Cell Line
Embryo cells
Embryos
Gene Expression Regulation, Developmental
Genes, Lethal - genetics
Genetic Vectors - genetics
Human Embryonic Stem Cells - metabolism
Humanities and Social Sciences
Humans
Mice
MicroRNAs - antagonists & inhibitors
MicroRNAs - genetics
miRNA
mRNA
multidisciplinary
Myocytes, Cardiac - metabolism
Myocytes, Cardiac - transplantation
Organogenesis - genetics
RNA, Messenger - genetics
Science
Science (multidisciplinary)
Stem cell transplantation
Stem cells
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Summary:Deriving cell populations from human embryonic stem cells (hESCs) for cell-based therapy is considered a promising strategy to achieve functional cells, yet its translation to clinical practice depends on achieving fully defined differentiated cells. In this work, we generated a miRNA-responsive lethal mRNA construct that selectively induces rapid apoptosis in hESCs by expressing a mutant (S184del) Bax variant. Insertion of miR-499 target sites in the construct enabled to enrich hESC-derived cardiomyocytes (CMs) in culture. A deterministic non-linear model was developed and validated with experimental data, to predict the outcome for each treatment cycle and the number of treatment cycle repetitions required to achieve completely purified cTNT-positive cells. The enriched hESC-CMs displayed physiological sarcomere orientation, functional calcium handling and after transplantation into SCID-NOD mice did not form teratomas. The modular miRNA responsive lethal mRNA construct could be employed in additional directed differentiation protocols, by adjusting the miRNA to the specific cells of choice.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-019-50899-2