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Isolation of Adipogenic and Fibro-Inflammatory Stromal Cell Subpopulations from Murine Intra-Abdominal Adipose Depots
Published on: August 16, 2020
Starvation and Inflammation Modulate Adipose Mesenchymal Stromal Cells' Molecular Signature
Simona Piccolo1, Giulio Grieco1, Caterina Visconte1
1Laboratorio di Biotecnologie Applicate all'Ortopedia, IRCCS Istituto Ortopedico Galeazzi, Via Cristina Belgioioso 173, 20157 Milano, Italy.
Abstract:
Mesenchymal stromal cells (MSCs) and their released factors (secretome) are intriguing options for regenerative medicine approaches based on the management of inflammation and tissue restoration, as in joint disorders like osteoarthritis (OA). Production strategy may modulate cells and secretome fingerprints, and for the latter, the effect of serum removal by starvation used in clinical-grade protocols has been underestimated. In this work, the effect of starvation on the molecular profile of interleukin 1 beta (IL1β)-primed adipose-derived MSCs (ASCs) was tested by assessing the expression level of 84 genes related to secreted factors and 84 genes involved in defining stemness potential. After validation at the protein level, the effect of starvation modulation in the secretomes was tested in a model of OA chondrocytes. IL1β priming in vitro led to an increase in inflammatory mediators' release and reduced anti-inflammatory potential on chondrocytes, features reversed by subsequent starvation. Therefore, when applying serum removal-based clinical-grade protocols for ASCs' secretome production, the effects of starvation must be carefully considered and investigated.
Insights
Serum starvation impacts the therapeutic potential of adipose-derived mesenchymal stromal cells (ASCs) secretomes. Starvation reversed inflammation induced by priming, highlighting its crucial role in regenerative medicine for osteoarthritis.
Area of Science:
- Regenerative Medicine
- Cell Biology
- Biochemistry
Background:
- Mesenchymal stromal cells (MSCs) and their secretomes show promise for treating inflammatory joint disorders like osteoarthritis (OA).
- Production methods significantly influence MSCs and their secretome characteristics.
- The impact of serum removal (starvation) in clinical-grade protocols for secretome production is often overlooked.
Purpose of the Study:
- To investigate the effect of serum starvation on the molecular profile of interleukin-1 beta (IL1β)-primed adipose-derived MSCs (ASCs).
- To assess how starvation affects the secretome's therapeutic potential in an osteoarthritis chondrocyte model.
Main Methods:
- ASCs were primed with IL1β, followed by serum starvation.
- Gene expression analysis was performed for 84 secreted factors and 84 stemness-related genes.
- Protein levels were validated, and secretome effects were tested on OA chondrocytes.
Main Results:
- IL1β priming increased inflammatory mediator release and decreased anti-inflammatory effects on chondrocytes.
- Subsequent serum starvation reversed these inflammatory effects induced by IL1β priming.
- Starvation significantly modulated the molecular profile and therapeutic function of ASC secretomes.
Conclusions:
- Serum starvation has a substantial impact on the molecular composition and functional properties of ASC secretomes.
- The effects of starvation must be carefully considered and investigated in clinical-grade protocols for ASC secretome production.
- Understanding starvation's role is critical for optimizing MSC-based therapies for osteoarthritis and other inflammatory conditions.
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