Application of human iPSC-derived macrophages in a miniaturized high-content-imaging-based efferocytosis assay

IF 2.7 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

SLAS Discovery Pub Date : 2023-06-01 DOI:10.1016/j.slasd.2023.04.002

Sarah Bitzer , Mozhgan Dehghan Harati , Karim C. El Kasmi , Daniela Schloesser , Julia Sauer , Heiko Olbrich , Michael Schuler , Florian Gantner , Ralf Heilker

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引用次数: 0

Abstract

Macrophages play a pivotal role in drug discovery due to their key regulatory functions in health and disease. Overcoming the limited availability and donor variability of human monocyte-derived macrophages (MDMs), human induced pluripotent stem cell (iPSC)-derived macrophages (IDMs) could provide a promising tool for both disease modeling and drug discovery. To access large numbers of model cells for medium- to high-throughput application purposes, an upscaled protocol was established for differentiation of iPSCs into progenitor cells and subsequent maturation into functional macrophages. These IDM cells resembled MDMs both with respect to surface marker expression and phago- as well as efferocytotic function. A statistically robust high-content-imaging assay was developed to quantify the efferocytosis rate of IDMs and MDMs allowing for measurements both in the 384- and 1536-well microplate format. Validating the applicability of the assay, inhibitors of spleen tyrosine kinase (Syk) were shown to modulate efferocytosis in IDMs and MDMs with comparable pharmacology. The miniaturized cellular assay with the upscaled provision of macrophages opens new routes to pharmaceutical drug discovery in the context of efferocytosis-modulating substances.

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人ipsc衍生巨噬细胞在小型化高含量成像的efferocytosis实验中的应用

巨噬细胞由于其在健康和疾病中的关键调节功能，在药物发现中发挥着关键作用。克服人类单核细胞衍生巨噬细胞（MDMs）的有限可用性和供体变异性，人类诱导多能干细胞（iPSC）衍生巨噬细胞（IDMs）可以为疾病建模和药物发现提供一种有前途的工具。为了获得大量的模型细胞用于中高通量应用，建立了一种将iPSC分化为祖细胞并随后成熟为功能性巨噬细胞的扩大方案。这些IDM细胞在表面标记物表达、吞噬和泡腾功能方面都类似于MDM。开发了一种统计上稳健的高含量成像测定法，以量化IDM和MDM的泡腾细胞增多率，从而允许以384孔和1536孔微孔板形式进行测量。为了验证该测定的适用性，脾脏酪氨酸激酶（Syk）抑制剂被证明可以通过类似的药理学调节IDMs和MDM的泡腾细胞增多症。在泡腾调节物质的背景下，巨噬细胞的小型化细胞测定为药物发现开辟了新的途径。

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来源期刊

SLAS Discovery Chemistry-Analytical Chemistry

CiteScore

7.00

自引率

3.20%

发文量

审稿时长

39 days

期刊介绍： Advancing Life Sciences R&D: SLAS Discovery reports how scientists develop and utilize novel technologies and/or approaches to provide and characterize chemical and biological tools to understand and treat human disease. SLAS Discovery is a peer-reviewed journal that publishes scientific reports that enable and improve target validation, evaluate current drug discovery technologies, provide novel research tools, and incorporate research approaches that enhance depth of knowledge and drug discovery success. SLAS Discovery emphasizes scientific and technical advances in target identification/validation (including chemical probes, RNA silencing, gene editing technologies); biomarker discovery; assay development; virtual, medium- or high-throughput screening (biochemical and biological, biophysical, phenotypic, toxicological, ADME); lead generation/optimization; chemical biology; and informatics (data analysis, image analysis, statistics, bio- and chemo-informatics). Review articles on target biology, new paradigms in drug discovery and advances in drug discovery technologies. SLAS Discovery is of particular interest to those involved in analytical chemistry, applied microbiology, automation, biochemistry, bioengineering, biomedical optics, biotechnology, bioinformatics, cell biology, DNA science and technology, genetics, information technology, medicinal chemistry, molecular biology, natural products chemistry, organic chemistry, pharmacology, spectroscopy, and toxicology. SLAS Discovery is a member of the Committee on Publication Ethics (COPE) and was published previously (1996-2016) as the Journal of Biomolecular Screening (JBS).