Comparison of two ASC-derived therapeutics in an in vitro OA model: secretome versus extracellular vesicles

Abstract Background In the last years, several clinical trials have proved the safety and efficacy of adipose-derived stem/stromal cells (ASC) in contrasting osteoarthritis (OA). Since ASC act mainly through paracrine mechanisms, their secretome (conditioned medium, CM) represents a promising therapeutic alternative. ASC-CM is a complex cocktail of proteins, nucleic acids, and lipids released as soluble factors and/or conveyed into extracellular vesicles (EV). Here, we investigate its therapeutic potential in an in vitro model of OA. Methods Human articular chondrocytes (CH) were induced towards an OA phenotype by 10 ng/ml TNFα in the presence of either ASC-CM or EV, both deriving from 5 × 105 cells, to evaluate the effect on hypertrophic, catabolic, and inflammatory markers. Results Given the same number of donor cells, our data reveal a higher therapeutic potential of ASC-CM compared to EV alone that was confirmed by its enrichment in chondroprotective factors among which TIMP-1 and -2 stand out. In details, only ASC-CM significantly decreased MMP activity (22% and 29% after 3 and 6 days) and PGE2 expression (up to 40% at day 6) boosted by the inflammatory cytokine. Conversely, both treatments down-modulated of ~ 30% the hypertrophic marker COL10A1. Conclusions These biological and molecular evidences of ASC-CM beneficial action on CH with an induced OA phenotype may lay the basis for its future clinical translation as a cell-free therapeutic in the management of OA.

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Extracellular Vesicles Are More Potent Than Adipose Mesenchymal Stromal Cells to Exert an Anti-Fibrotic Effect in an In Vitro Model of Systemic Sclerosis

International Journal of Molecular Sciences ◽

10.3390/ijms22136837 ◽

2021 ◽

Vol 22 (13) ◽

pp. 6837

Author(s):

Pauline Rozier ◽

Marie Maumus ◽

Claire Bony ◽

Alexandre Thibault Jacques Maria ◽

Florence Sabatier ◽

...

Keyword(s):

Systemic Sclerosis ◽

Extracellular Vesicles ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

In Vitro Model ◽

Specific Treatment ◽

Complex Disorder ◽

Molecular Features ◽

Vitro Model

Systemic sclerosis (SSc) is a complex disorder resulting from dysregulated interactions between the three main pathophysiological axes: fibrosis, immune dysfunction, and vasculopathy, with no specific treatment available to date. Adipose tissue-derived mesenchymal stromal cells (ASCs) and their extracellular vesicles (EVs) have proved efficacy in pre-clinical murine models of SSc. However, their precise action mechanism is still not fully understood. Because of the lack of availability of fibroblasts isolated from SSc patients (SSc-Fb), our aim was to determine whether a TGFβ1-induced model of human myofibroblasts (Tβ-Fb) could reproduce the characteristics of SSc-Fb and be used to evaluate the anti-fibrotic function of ASCs and their EVs. We found out that Tβ-Fb displayed the main morphological and molecular features of SSc-Fb, including the enlarged hypertrophic morphology and expression of several markers associated with the myofibroblastic phenotype. Using this model, we showed that ASCs were able to regulate the expression of most myofibroblastic markers on Tβ-Fb and SSc-Fb, but only when pre-stimulated with TGFβ1. Of interest, ASC-derived EVs were more effective than parental cells for improving the myofibroblastic phenotype. In conclusion, we provided evidence that Tβ-Fb are a relevant model to mimic the main characteristics of SSc fibroblasts and investigate the mechanism of action of ASCs. We further reported that ASC-EVs are more effective than parental cells suggesting that the TGFβ1-induced pro-fibrotic environment may alter the function of ASCs.

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Activation by IL-1 of bovine articular chondrocytes in culture within a 3D collagen-based scaffold. An in vitro model to address the effect of compounds with therapeutic potential in osteoarthritis

Osteoarthritis and Cartilage ◽

10.1016/j.joca.2006.01.008 ◽

2006 ◽

Vol 14 (7) ◽

pp. 631-640 ◽

Cited By ~ 34

Author(s):

D. Cortial ◽

J. Gouttenoire ◽

C.F. Rousseau ◽

M.-C. Ronzière ◽

N. Piccardi ◽

...

Keyword(s):

In Vitro Model ◽

Articular Chondrocytes ◽

Therapeutic Potential ◽

3D Collagen ◽

Bovine Articular Chondrocytes ◽

Vitro Model

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Faculty Opinions recommendation of Transcriptional response of human articular chondrocytes treated with fibronectin fragments: an in vitro model of the osteoarthritis phenotype.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.739123584.793581017 ◽

2020 ◽

Author(s):

Peter Torzilli

Keyword(s):

In Vitro Model ◽

Articular Chondrocytes ◽

Transcriptional Response ◽

Human Articular Chondrocytes ◽

Fibronectin Fragments ◽

Vitro Model

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Transcriptional response of human articular chondrocytes treated with fibronectin fragments: an in vitro model of the osteoarthritis phenotype

Osteoarthritis and Cartilage ◽

10.1016/j.joca.2020.09.006 ◽

2020 ◽

Author(s):

K.S.M. Reed ◽

V. Ulici ◽

C. Kim ◽

S. Chubinskaya ◽

R.F. Loeser ◽

...

Keyword(s):

In Vitro Model ◽

Articular Chondrocytes ◽

Transcriptional Response ◽

Human Articular Chondrocytes ◽

Fibronectin Fragments ◽

Vitro Model

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Exosome derived from murine adipose-derived stromal cells: Neuroprotective effect on in vitro model of amyotrophic lateral sclerosis

Experimental Cell Research ◽

10.1016/j.yexcr.2015.12.009 ◽

2016 ◽

Vol 340 (1) ◽

pp. 150-158 ◽

Cited By ~ 61

Author(s):

Roberta Bonafede ◽

Ilaria Scambi ◽

Daniele Peroni ◽

Valentina Potrich ◽

Federico Boschi ◽

...

Keyword(s):

Amyotrophic Lateral Sclerosis ◽

Stromal Cells ◽

In Vitro Model ◽

Neuroprotective Effect ◽

Adipose Derived Stromal Cells ◽

Vitro Model ◽

Lateral Sclerosis

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Growth factor and cytokine expression of human mesenchymal stromal cells is not altered in an in vitro model of tissue damage

Cytotherapy ◽

10.3109/14653249.2010.501789 ◽

2010 ◽

Vol 12 (7) ◽

pp. 870-880 ◽

Cited By ~ 13

Author(s):

Katrin Montzka ◽

Tobias Führmann ◽

Jochen Müller-Ehmsen ◽

Michael Wöltje ◽

Gary A. Brook

Keyword(s):

Growth Factor ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Tissue Damage ◽

In Vitro Model ◽

Cytokine Expression ◽

Human Mesenchymal Stromal Cells ◽

Vitro Model

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SU-FF-J-104: In Vitro Model to Study the Biological Effect of Dose Gradients On a Cell Culture System

Medical Physics ◽

10.1118/1.1997650 ◽

2005 ◽

Vol 32 (6Part6) ◽

pp. 1944-1944 ◽

Cited By ~ 2

Author(s):

R Bromley ◽

L Oliver ◽

R Harvie ◽

R Davey

Keyword(s):

Cell Culture ◽

Biological Effect ◽

Culture System ◽

In Vitro Model ◽

Cell Culture System ◽

A Cell ◽

Vitro Model

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An in vitro model of erythroid egress in bone marrow

Blood ◽

10.1182/blood.v68.1.250.250 ◽

1986 ◽

Vol 68 (1) ◽

pp. 250-257 ◽

Cited By ~ 7

Author(s):

RE Waugh ◽

M Sassi

Keyword(s):

Bone Marrow ◽

In Vitro Model ◽

Physical Parameters ◽

In Vitro System ◽

Wafer Thickness ◽

Pore Cells ◽

A Cell ◽

Vitro Model ◽

Passage Times

Abstract An in vitro system has been developed that mimics the passage of erythrocytes from the bone marrow to the circulation. Bone marrow egress and its proper regulation are vital physiologic processes. However, because of the inaccessibility of the marrow, it is difficult to evaluate the various factors important in controlling these processes or even to define the precise mechanism by which egress occurs. The in vitro system has been designed to evaluate the importance of different physical parameters in regulating egress. It consists of a thin silicon wafer (thickness approximately equal to 1.0 micron) cemented over the tip of a large (15.0 micron ID) micropipette. The wafer contains a single circular pore. Cells were observed under the microscope as they passed through the pore under controlled pressures. The rate and duration of passage were obtained from videorecordings of the experiment. The measured passage times agreed well with the predictions of a simple analytical model of a cell passing through a thin aperture. The experimental results confirm the conclusion reached from the analysis that the pressures needed to drive a cell through the pore are well within the physiologic range, and the time needed to complete egress is typically less than 1.0 seconds. These results support the hypothesis that erythrocyte egress may be driven by a hydrostatic pressure difference across the pore.

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