DATE: June 7, 2017
TIME: 9:00AM PDT, 12:00PM EDT
Pluripotent stem cells (PSCs) offer an unlimited source of human cardiovascular cells for research and the development of cardiac regeneration therapies. The development of highly efficient cardiac-directed differentiation methods makes it possible to generate large numbers of cardiomyocytes (hPSC-CMs). Due to varying differentiation efficiencies, further enrichment of CM populations for downstream applications is essential. Recently, a CM-specific cell surface marker called SIRPa (signal-regulatory protein alpha, also termed CD172a) was reported to be a useful tool for flow sorting of human stem cell–derived CMs. However, our expression analysis revealed that SIRPa only labels a subpopulation of CMs indicated by cardiac Troponin T (cTnT) expression. Moreover, SIRPa is also expressed on a sub population of non-CMs, hence making SIRa an inadequate marker to enrich PSC-derived CMs.
To circumvent the disadvantages of SIRPa for CM enrichment, we further evaluated the novel PSC-Derived Cardiomyocyte Isolation Kit, human, which utilizes different surface markers than SIRPa for CM enrichment. The kit provides two different strategies for CM enrichment which can be applied depending on the initial differentiation efficiency. For differentiation cultures with a CM content above 50% a single separation step involving depletion of non-CMs is sufficient to achieve purities >90%. For low differentiation efficiencies below 50% a second enrichment step, utilizing a novel, highly specific CM marker, can be added to further increase purity. To assess and compare the kit performance to SIRPa-dependent enrichment, the CMs were analyzed phenotypically and functionally after the cell separation step. In this webinar the process for hiPSC derivation, cardiac directed differentiation to hPSC-CMs, enrichment of hPSC-CMs and subsequent formation of 2D monolayers of electrically connected cells will be reviewed.
The content of the following presentation by Todd Heron, has not been prepared by, interpreted by or reviewed by Miltenyi Biotec Inc. The views and opinions expressed in the following presentation are solely those of the investigator. All Miltenyi Biotec Inc.’s products discussed herein are for Research Use Only unless otherwise specified.