15-02-2018 дата публикации
Номер: US20180042969A1
Принадлежит:
The present disclosure provides compositions and methods for the reprogramming of cells such as fibroblasts into cardiomyocytes. The invention provided herein features a chemically defined media and methods of reprogramming cells to increase cardiac gene and protein expression in cardiac fibroblasts and other fibroblasts, e.g. dermal fibroblasts. The media and methods also enhance miR-combo mediated cardiac reprogramming of fibroblasts to cardiomyocytes. Thus, the invention encompasses a chemically defined reprogramming media comprising a base tissue culture media, insulin-transferrin-selenium (ITS) or ascorbic acid in a somatic cell-reprogramming, e.g., fibroblast-to-cardiomyocyte-reprogramming, amount. 1. A chemically defined reprogramming media comprising a base tissue culture media , insulin-transferrin-selenium or ascorbic acid in a somatic cell-reprogramming amount.2. The media of claim 1 , wherein said media further comprises bovine serum albumin (BSA) or claim 1 , L-glutamine.3. The media of claim 1 , wherein said media comprises insulin-transferrin-selenium claim 1 , said insulin being present in an amount of 10 nanomolar to 10 micromolar claim 1 , said transferrin being present in an amount of 0.002 to lgram per liter claim 1 , and said selenium being present in an amount of 1-100 μg per liter.4. The media of claim 1 , wherein said media comprises 0.2 mM to 20 mM L-glutamine.5. The media claim 1 , wherein said media comprises 50 μM to 50 millimolar ascorbic acid.6. The media of claim 1 , further comprising a reprogramming efficiency-enhancing molecule.7. The media of claim 6 , wherein said molecule is one or more molecules selected from the group consisting of valproic acid claim 6 , bone morphogenetic protein 4 (BMP4) claim 6 , JAK inhibitor 1 claim 6 , RG108 claim 6 , R(+) Bay K 8644 claim 6 , PS48 claim 6 , and A83-01.8. A method of reprogramming a cell into a fibroblast comprising contacting the cell with the media according to for a sufficient amount ...
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