期刊
JOURNAL OF LIPID RESEARCH
卷 59, 期 7, 页码 1301-1310出版社
ELSEVIER
DOI: 10.1194/jlr.D085217
关键词
adipogenesis; adipocyte; label-free
资金
- Swiss National Science Foundation [PP00P3_144857]
- Fondation Pierre Mercier pour la Science
- Fondation Dr Henri Dubois-Ferriere Dinu Lipatti
- Fundacion Josep Carreras Contra la Leucemia
- European Hematology Association
- Swiss National Science Foundation (SNF) [PP00P3_144857] Funding Source: Swiss National Science Foundation (SNF)
In vitro differentiating adipocytes are sensitive to liquid manipulations and have the tendency to float. Assessing adipocyte differentiation using current microscopy techniques involves cell staining and washing, while using flow cytometry involves cell retrieval in suspension. These methods induce biases, are difficult to reproduce, and involve tedious optimizations. In this study, we present digital holographic microscopy (DHM) as a label-free, nonperturbing means to quantify lipid droplets in differentiating adipocytes in a robust medium- to high-throughput manner. Taking advantage of the high refractive index of lipid droplets, DHM can assess the production of intracellular lipid droplets by differences in phase shift in a quantitative manner. Adipocytic differentiation, combined with other morphological features including cell confluence and cell death, was tracked over 6 days in live OP9 mesenchymal stromal cells. We compared DHM with other currently available methods of lipid droplet quantification and demonstrated its robustness with modulators of adipocytic differentiation in a dose-responsive manner. This study suggests DHM as a novel marker-free nonperturbing method to study lipid droplet accumulation and may be envisioned for drug screens and mechanistic studies on adipocytic differentiation.
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