ArticleProceedings of the National Academy of Sciences of the United States of America2016
In situ characterization of the mTORC1 during adipogenesis of human adult stem cells on chip.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed.
- Alternative Methods as Tools for Obesity Research: In Vitro and In Silico Approaches.Life (Basel, Switzerland) · 2022Review
- Lysosome Inhibition Reduces Basal and Nutrient-Induced Fat Accumulation inMolecules and cells · 2022Article
- Adipose microtissue-on-chip: a 3D cell culture platform for differentiation, stimulation, and proteomic analysis of human adipocytes.Lab on a chip · 2022Article
- Strategies for improving adipose-derived stem cells for tissue regeneration.Burns & trauma · 2022Review
- Probing Insulin Sensitivity with Metabolically Competent Human Stem Cell-Derived White Adipose Tissue Microphysiological Systems.Small (Weinheim an der Bergstrasse, Germany) · 2022Article
- Highly parallelized human embryonic stem cell differentiation to cardiac mesoderm in nanoliter chambers on a microfluidic chip.Biomedical microdevices · 2021Article
- A 3D human adipose tissue model within a microfluidic device.Lab on a chip · 2021Article
- Fat-On-A-Chip Models for Research and Discovery in Obesity and Its Metabolic Comorbidities.Tissue engineering. Part B, Reviews · 2020Review
- Fattening chips: hypertrophy, feeding, and fasting of human white adipocytesLab on a chip · 2020Article
- Review
- Metallic Nanoparticle-Based Optical Cell Chip for Nondestructive Monitoring of Intra/Extracellular Signals.Pharmaceutics · 2020Review
- Modular operation of microfluidic chips for highly parallelized cell culture and liquid dosing via a fluidic circuit board.Microsystems & nanoengineering · 2020Article
- Arrayed isoelectric focusing using photopatterned multi-domain hydrogels.Electrophoresis · 2018Article
- Microfluidic systems for studying dynamic function of adipocytes and adipose tissue.Analytical and bioanalytical chemistry · 2018Review
- Multiplexed Point-of-Care Testing - xPOCT.Trends in biotechnology · 2017Review
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Authors and funding
8 authors.
Funding
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Abstract
Mammalian target of rapamycin (mTOR) is a central kinase integrating nutrient, energy, and metabolite signals. The kinase forms two distinct complexes: mTORC1 and mTORC2. mTORC1 plays an essential but undefined regulatory function for regeneration of adipose tissue. Analysis of mTOR in general is hampered by the complexity of regulatory mechanisms, including protein interactions and/or phosphorylation, in an ever-changing cellular microenvironment. Here, we developed a microfluidic large-scale integration chip platform for culturing and differentiating human adipose-derived stem cells (hASCs) in 128 separated microchambers under standardized nutrient conditions over 3 wk. The progression of the stem cell differentiation was measured by determining the lipid accumulation rates in hASC cultures. For in situ protein analytics, we developed a multiplex in situ proximity ligation assay (mPLA) that can detect mTOR in its two complexes selectively in single cells and implemented it on the same chip. With this combined technology, it was possible to reveal that the mTORC1 is regulated in its abundance, phosphorylation state, and localization in coordination with lysosomes during adipogenesis. High-content image analysis and parameterization of the in situ PLA signals in over 1 million cells cultured on four individual chips showed that mTORC1 and lysosomes are temporally and spatially coordinated but not in its composition during adipogenesis.
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