A call to adopt a 'fit-for-purpose' approach to antibody validation for flow cytometry analyses of stem cell models and beyond.

A call to adopt a 'fit-for-purpose' approach to antibody validation for flow cytometry analyses of stem cell models and beyond. Am J Physiol Heart Circ Physiol. 2019 Sep 27;: Authors: Waas M, Gundry RL Abstract Human pluripotent stem cell-derived cardiomyocytes (hPSC-CM) can be exploited as models for a wide range of research applications and numerous protocols for generating hPSC-CM have been described. However, it is currently not possible to direct differentiation to a single, homogeneous endpoint, and the resulting heterogeneity may be variable among laboratories, cell lines, and protocols. Consequently, the ability to assess phenotypic heterogeneity of the cell population is critical to the interpretation, repeatability, and reproduction of hPSC-CM studies. While flow cytometry is well-suited for this purpose, a review of published literature reveals there is currently no consensus regarding which marker, antibody, or protocol is best suited to enable comparisons of hPSC-CM culture heterogeneity. Moreover, the lack of available experimental detail, combined with the variability in the approaches used for hPSC-CM evaluation, makes it challenging to reproduce, interpret, and compare published data. Consequently, this Perspective calls for an alignment of the way researchers approach the routine use and documentation of the antibodies and controls used during flow cytometry-based assessment of hPSC-CM cultures. We advocate for the ...
Source: American Journal of Physiology. Heart and Circulatory Physiology - Category: Physiology Authors: Tags: Am J Physiol Heart Circ Physiol Source Type: research