Mass spectrometry-based proteomics of 3D cell culture: A useful tool to validate culture of spheroids and organoids.

Autor:	Avelino TM; National Center of Research in Energy and Materials (CNPEM), National Laboratory of Bioscience (LNBio); State University of Campinas (UNICAMP), Department of Pharmacology Science., García-Arévalo M; National Center of Research in Energy and Materials (CNPEM), National Laboratory of Bioscience (LNBio)., Torres FR; National Center of Research in Energy and Materials (CNPEM), National Laboratory of Bioscience (LNBio)., Goncalves Dias MM; National Center of Research in Energy and Materials (CNPEM), National Laboratory of Bioscience (LNBio); State University of Campinas (UNICAMP), Department of Molecular and Functional Biology., Domingues RR; National Center of Research in Energy and Materials (CNPEM), National Laboratory of Bioscience (LNBio)., de Carvalho M; National Center of Research in Energy and Materials (CNPEM), National Laboratory of Bioscience (LNBio)., Fonseca MC; National Center of Research in Energy and Materials (CNPEM), National Laboratory of Bioscience (LNBio)., Rodrigues VKT; National Center of Research in Energy and Materials (CNPEM), National Laboratory of Bioscience (LNBio)., Leme AFP; National Center of Research in Energy and Materials (CNPEM), National Laboratory of Bioscience (LNBio)., Figueira ACM; National Center of Research in Energy and Materials (CNPEM), National Laboratory of Bioscience (LNBio); State University of Campinas (UNICAMP), Department of Pharmacology Science; State University of Campinas (UNICAMP), Department of Molecular and Functional Biology. Electronic address: ana.figueira@lnbio.cnpem.br.
Jazyk:	angličtina
Zdroj:	SLAS discovery : advancing life sciences R & D [SLAS Discov] 2022 Apr; Vol. 27 (3), pp. 167-174. Date of Electronic Publication: 2021 Oct 27.
DOI:	10.1016/j.slasd.2021.10.013
Abstrakt:	Worldwide obesity, defined as abnormal or excessive fat accumulation that may result in different comorbidities, is considered a pandemic condition that has nearly tripled in the last 45 years. Most studies on obesity use animal models or adipocyte monolayer cell culture to investigate adipose tissue. However, besides monolayer cell culture approaches do not fully recapitulate the physiology of living organisms, there is a growing need to reduce or replace animals in research. In this context, the development of 3D self-organized structures has provided models that better reproduce the in vitro aspects of the in vivo physiology in comparison to traditional monolayer cell culture. Besides, recent advances in omics technologies have allowed us to characterize these cultures at the proteome, metabolome, transcription factor, DNA-binding and transcriptomic levels. These two combined approaches, 3D culture and omics, have provided more realistic data about determined conditions. Thereby, here we focused on the development of an obesity study pipeline including proteomic analysis to validate adipocyte-derived spheroids. Through the combination of collected mass spectrometry data from differentiated 3T3-L1 spheroids and from murine white adipose tissue (WAT), we identified 1732 proteins in both samples. By using a comprehensive proteomic analysis, we observed that the in vitro 3D culture of differentiated adipocytes shares important molecular pathways with the WAT, including expression of proteins involved in central metabolic process of the adipose tissue. Together, our results show a combination of an orthogonal method and an image-based analysis that constitutes a useful pipeline to be applied in 3D adipocyte culture. Competing Interests: Declaration of Competing Interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as potential conflict of interest. This article is being reproduced in print post-publication in a sponsored print collection for distribution. The company sponsoring the print collection was not involved in the editorial selection or review of this article. (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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