Suppression of Pro-fibrotic Signaling Potentiates Factor-mediated Reprogramming of Mouse Embryonic Fibroblasts into Induced Cardiomyocytes.

摘要: Trans-differentiation of one somatic cell type into another has enormous potential to model and treat human diseases. Previous studies have shown that mouse embryonic, dermal, cardiac fibroblasts can be reprogrammed functional induced-cardiomyocyte-like cells (iCMs) through overexpression cardiogenic transcription factors including GATA4, Hand2, Mef2c, Tbx5 both in vitro vivo. However, these previous relatively low efficiency. In order restore heart function following injury, mechanisms governing reprogramming must elucidated increase efficiency maturation iCMs. We previously demonstrated inhibition pro-fibrotic signaling dramatically increases Here, we detail methods achieve a up 60%. Furthermore, describe several flow cytometry, immunofluorescent imaging, calcium imaging quantify fibroblasts. Using the protocol detailed here, mechanistic undertaken determine positive negative regulators reprogramming. These may identify pathways targeted promote maturation, which could lead novel therapies disease.