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Noninvasive Continuous Monitoring of Adipocyte Differentiation: From Macro to Micro Scales

Published online by Cambridge University Press:  04 February 2019

Maayan Lustig Open the ORCID record for Maayan Lustig [Opens in a new window] ,
Qingling Feng ,
Yohan Payan ,
Amit Gefen  and
Dafna Benayahu
Maayan Lustig
Affiliation:
Department of Biomedical Engineering, Faculty of Engineering, Tel Aviv University, Tel Aviv 6997801, Israel
Qingling Feng
Affiliation:
Key Laboratory of Advanced Materials of Ministry of Education of China, School of Materials Science and Engineering, Tsinghua University, Beijing 100084, China
Yohan Payan
Affiliation:
CNRS, Grenoble INP, TIMC-IMAG, University of Grenoble Alpes, Grenoble F-38000, France
Amit Gefen
Affiliation:
Department of Biomedical Engineering, Faculty of Engineering, Tel Aviv University, Tel Aviv 6997801, Israel
Dafna Benayahu*
Affiliation:
Department of Cell and Developmental Biology, Sackler School of Medicine, Tel Aviv University, Tel Aviv 6997801, Israel
*
*Author for correspondence: Dafna Benayahu, E-mail: dafnab@tauex.tau.ac.il
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Abstract

3T3-L1 cells serve as model systems for studying adipogenesis and research of adipose tissue-related diseases, e.g. obesity and diabetes. Here, we present two novel and complementary nondestructive methods for adipogenesis analysis of living cells which facilitate continuous monitoring of the same culture over extended periods of time, and are applied in parallel at the macro- and micro-scales. At the macro-scale, we developed visual differences mapping (VDM), a novel method which allows to determine level of adipogenesis (LOA)—a numerical index which quantitatively describes the extent of differentiation in the whole culture, and percentage area populated by adipocytes (PAPBA) across a whole culture, based on the apparent morphological differences between preadipocytes and adipocytes. At the micro-scale, we developed an improved version of our previously published image-processing algorithm, which now provides data regarding single-cell morphology and lipid contents. Both methods were applied here synergistically for measuring differentiation levels in cultures over multiple weeks. VDM revealed that the mean LOA value reached 1.11 ± 0.06 and the mean PAPBA value reached >60%. Micro-scale analysis revealed that during differentiation, the cells transformed from a fibroblast-like shape to a circular shape with a build-up of lipid droplets. We predict a vast potential for implementation of these methods in adipose-related pharmacological research, such as in metabolic-syndrome studies.

Keywords

adipogenesismorphologylipid dropletsimage processingmapping
Type
Biological Science Applications
Information
Microscopy and Microanalysis , Volume 25 , Issue 1 , February 2019 , pp. 119 - 128
Copyright
Copyright © Microscopy Society of America 2019 

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