JUN 07, 2017 9:00 AM PDT

WEBINAR: Generation of purified human iPSC derived cardiomyocytes using clinically relevant workflow

Sponsored by: Miltenyi Biotec
Speaker
  • Director of the Frankel Cardiovascular Center's Cardiovascular Regeneration Core Laboratory and Assistant Research Professor at the University of Michigan Center for Arrhythmia Research
    Biography
      Dr. Herron is the Director of the Frankel Cardiovascular Center's Cardiovascular Regeneration Core Laboratory. He is on the faculty in the University of Michigan Medical School and has appointments in the Department of Internal Medicine and Molecular & Integrative Physiology as Associate Research Scientist. His research is focused on the complex interplay between cardiac electrical excitation and contractile force generation-a process known classically as excitation-contraction coupling. Excitation-contraction coupling is highly regulated in the mammalian heart and slight modifications of this process occur on a beat-to-beat basis and any dysfunction can lead to fatal cardiac arrhythmias and heart failure. In this research Dr. Herron has utilized transgenic animals, surgical models of heart failure and more recently human induced pluripotent stem cells (hiPSCs). hiPSC technology has revolutionized the biomedical research arena and using highly efficient differentiation techniques it is possible now to create an unlimited supply of patient specific cardiovascular cells for study and for future therapy development. Dr. Herron's lab was among the first to utilize hiPSC-CM monolayers to study human cardiac arrhythmia mechanisms in vitro. Current research is focused on developing patient specific cardiomyopathy disease models in vitro using hiPSC-CM technology as well as development of high throughput screening platforms to test for compound effects on human cardiac function. A primary goal of the Core Laboratory directed by Dr. Herron is to work towards generation of clinically relevant hiPSCs and hiPSC-CMs by using Xeno-free media and non-genetic modification based purification of derived cells. In this endeavor his lab relies on clinically relevant reagents from Miltenyi Biotec. By using magnetic activated cell sorting his lab is now able to generate large numbers of highly enriched hiPSC-CMs for basic science research purposes.

    Abstract

    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.

    Learning Objectives:

    • Review human induced pluripotent stem cell derivation, cardiac directed differentiation to human pluripotent stem cell cardiomyocytes (hPSC-CMs), enrichment of hPSC-CMs and subsequent formation of 2D monolayers of electrically connected cells.
    • Generation of purified human induced pluripotent stem cell derived cardiomyocyte

    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.


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