Label-Free, Non-Genetic Identification and Sorting of Human Pluripotent Stem Cell Derived Cardiomyocytes
Identifying stem cell derived cardiomyocytesSorting for pure populations of stem cell derived cardiomyocytesRemoving pluripotent stem cells from a cardiomyocyte populationAssessing maturity of cardiomyocytesDerivation of clinical grade cardiomyocytesLabel-free (xeno-free)Non-genetic (vector-free)Detection of very immature cardiomyocytesDiscrimination of cells at different stages of maturation
UC Davis researchers from the NSF Center for Biophotonics and UC Davis Health System have developed a method of identifying and sorting cardiomyocytes derived from human pluripotent stem cells. This method, based on second harmonic generation (SHG) - a nonlinear optical technique, does not require genetic modification of the cell or any exogenous labels to be used, which makes this an attractive technique for obtaining pure populations of cardiomyocytes under xeno- and vector- free conditions most appropriate for clinical and therapeutic use, as well for tissue engineering and drug discovery applications. There are currently no established methods for sorting pur populations of stem cell derived cardiomyocytes. Methods that use fluorescent reporters require the introduction of a reporter vector and result in genetically modified cells, reducing their utility for clinical applications. Other fluorescent-based staining methods have shown to be only applicable for selecting very mature cardiomyocytes. Surface marker based methods require exposing human cells to products of animal origin, which may increase the risk of non-human pathogen transmission and render the cells unsuitable for clinical use. Second harmonic generation (SHG) is a laser-based technique that identifies stem cell derived cardiomyocytes based on the direct detection of myosin bundles, which generates a unique second harmonic signal when excited by intense laser pulses. This signal is specific to the cardiomyocyte phenotype and is absent from undifferentiated stem cells and other non-cardiomyocyte cells that are found in the population following the directed differentiation of stem cells to the cardiac lineage. SHG is able to discriminate cardionmyocytes at different stages of maturation/development, and can detect very immature cells. When integrated into a flow cytometric configuration, non-invasive sorting for pure populations of stem cell derived cardiomyocytes is feasible.
9879224
Additional Technologies by these Inventors Tech ID/UC Case 22045/2010-609-0 Related Cases 2010-609-0
美國
