Utilizing Glucose Depletion to Purify Cardiomyocytes

Abstract

Cardiovascular disease (CVD) is a leading cause of death in the developed world affecting an estimated 83.6 million American adults. Cardiac cell types derived from human pluripotent stem cells (hPSCs), including human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs), are the primary cellular building blocks being investigated for treatment and repair of cardiac damage. Many of these techniques result in cell populations that are mixed with undifferentiated cells and non-cardiomyocytes. A variety of protocols have been established to increase the efficiency of hPSC cardiac differentiation, but the generation of purified populations has been a progress limitation A purified population of CMs would provide tissue engineers and researchers alike with a control for experiments regarding cardiovascular tissue regeneration of any form. Recently the metabolic biochemical difference in glucose and lactate uptake between cardiomyocytes and non-cardiomyocytes has been targeted as a method for purification. By taking advantage of this unique metabolic property of cardiomyocytes we can develop a noninvasive, inexpensive and efficient method to create a homogenous population. By applying glucose depletion protocol at identified time point, Day 10 and Day 20 post-differentiation we were able to provide population purities of up to 95%.