Addressing variability in iPSC-derived models of human disease: guidelines to promote reproducibility.

Viola Volpato,Caleb Webber

doi:10.1242/dmm.042317

Abstract

ABSTRACTInduced pluripotent stem cell (iPSC) technologies have provided in vitro models of inaccessible human cell types, yielding new insights into disease mechanisms especially for neurological disorders. However, without due consideration, the thousands of new human iPSC lines generated in the past decade will inevitably affect the reproducibility of iPSC-based experiments. Differences between donor individuals, genetic stability and experimental variability contribute to iPSC model variation by impacting differentiation potency, cellular heterogeneity, morphology, and transcript and protein abundance. Such effects will confound reproducible disease modelling in the absence of appropriate strategies. In this Review, we explore the causes and effects of iPSC heterogeneity, and propose approaches to detect and account for experimental variation between studies, or even exploit it for deeper biological insight.

Highlights

Since their invention just over a decade ago, induced pluripotent stem cell-based models have established a new field in disease modelling, especially for neurological disorders for which inadequate preclinical animal models and poor access to human primary tissue are limiting progress
Fly or worm models are usually generated within a small number of well-studied genetic backgrounds, thousands of new human induced pluripotent stem cell (iPSC) lines have been generated in the UK alone in the past 5 years, each influenced by its unique genetic background (Box 1) and with the vast majority individually receiving very little study
The process of induced reprogramming (Box 1) is based on erasing the existing epigenetic state of the cell of origin (Bilic et al, 2012; Medvedev et al, 2012), the tissue from which the iPSCs were derived and the retention of specific DNA methylation marks can determine the propensity of a line to differentiate into different cell types

Summary

Introduction

Since their invention just over a decade ago, induced pluripotent stem cell (iPSC; see Glossary, Box 1)-based models have established a new field in disease modelling, especially for neurological disorders for which inadequate preclinical animal models and poor access to human primary tissue are limiting progress. These approaches can ameliorate some of the drawbacks of iPSC-based models such as reduced cell maturity, incomplete disease phenotypes and line-to-line variation (Ghaffari et al, 2018), these limitations still need to be effectively addressed to be able to work with patient-derived cells in a high-standard, reproducible and controlled environment.

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Conclusion