Reversibly immortalizing cardiomyocytes to enable cell expansion in cultures
Lead Inventors: Gordana Vunjak-Novakovic, Ph.D.; Yue ZhangProblem or Unmet Need:Tissue engineering a clinically useful patch of cardiac tissue to treat the massive loss of cardiomyocytes that typically occurs during myocardial infarctions requires a large number of cells in order to ensure functional cell coupling and signaling. Although the use of induced pluripotent cells may offer a plentiful source of autologous cardiomyocytes, it still remains to be seen how much phenotypic stabilization needs to be done in order to avoid the risk of tumor formation. Consequently, there is a need for alternative techniques for obtaining large numbers of cardiomyocytes that can be used for cardiac tissue engineering.This technology is a method for controlled expansion of primary cardiomyocytes by reversible genetic engineering. The method involves the immortalization of cardiomyocytes by introducing the immortalizing genes large T antigen (TAg) or Bmi-1 combined with the human telomerase reverse transcriptase (hTERT) gene to the cells via lentiviral vectors, expanding the immortalized cells, and returning the expanded cells to their differentiated state by removing the immortalization genes with transient expression of Cre recombinase delivered by adenoviral transfection.
This reversible immortalization strategy can temporarily immortalize terminally differentiated and non-proliferating primary cells, and then return them to the "normal" differentiated state by removing the inserted gene by Cre-loxP site recombination. Expanding a patient's own cardiomyocytes via reversible immortalization could avoid the possible immunogenic or tumorigenic problems that may be caused by the use of exogenous or induced pluripotent stem cells.
The technology could be used to expand large numbers of autologous cardiomyocytes from a small biopsy sample for cell therapy. The cardiomyocytes derived by this method could be used as a cellular model with which to study cardiogenesis.
This technology is a method for controlled expansion of primary cardiomyocytes by reversible genetic engineering. The method involves the immortalization of cardiomyocytes by introducing the immortalizing genes large T antigen (TAg) or Bmi-1 combined...
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