Reprogramming of Human Urine Stem Cells into Cardiomyocytes via a Small Molecule Cocktail in Xeno-free conditions
Reprogramming autologous somatic cells into alternative lineages holds significant promise as a crucial cell source for regenerative therapy. Among various sources, human urine stem cells (hUSCs) have emerged as a highly promising candidate for reprogramming due to their ease of collection and high transdifferentiation potential. In this study, we present the first evidence for cardiac regeneration by directly converting hUSCs into functional cardiomyocytes under xeno-free conditions using a cocktail of 15 small molecules. This innovative protocol ensures a safe, animal-free reprogramming environment, enhancing its clinical applicability. The resulting chemically induced human cardiomyocyte-like cells (hCiCMs) exhibit several key characteristics of native cardiomyocytes, including the expression of cardiomyocyte-specific markers, display sarcomeric organization, and exhibit typical cardiac calcium flux and electrophysiological properties. Notably, transplantation of hCiCMs into the infarcted hearts of mice led to substantial improvements in cardiac function, characterized by enhanced contractility and reduced infarct size. Our findings validate the efficacy of the xeno-free differentiation protocol and highlight the potential of hUSC-derived hCiCMs as a viable and effective source for regenerative therapies in myocardial infarction. The use of hUSCs provides a readily accessible, autologous cell source, thereby paving the way for personalized and safer regenerative treatments for cardiac diseases. Overall design: Digest the cells on day 60 of reprogramming, and then perform single-cell sequencing.