Controlled Aggregation Enhances Immunomodulatory Potential of Mesenchymal Stromal Cell Aggregates

Xie, Angela W.; Zacharias, Nicholas; Binder, Bernard Y. K.; Murphy, William L.

doi:10.1002/sctm.19-0414

Cited by 19 publications

(14 citation statements)

References 89 publications

(161 reference statements)

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“…CXCR4 (involved in MSC homing in the body) and IL-24 (anti-cancer factor) expressions have shown to be increased in MSC spheroids ( Frith et al, 2010 ). When cultivated as 3D aggregates or spheroids, MSCs display increased angiogenic, anti-inflammatory, and immunomodulatory effects as well as improved stemness and survival rates after transplantation ( Xie et al, 2021 ). It has been suggested that cultivation of MSC spheroids could modulate their differentiation, proliferation and paracrine effects ( Egger et al, 2018 ).…”

Section: Introductionmentioning

confidence: 99%

Effect of 2D and 3D Culture Microenvironments on Mesenchymal Stem Cell-Derived Extracellular Vesicles Potencies

Kusuma

Zhu

et al. 2022

Front. Cell Dev. Biol.

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Therapeutic benefits of mesenchymal stem cells (MSCs) are now widely believed to come from their paracrine signalling, i.e. secreted factors such as cytokines, chemokines, and extracellular vesicles (EVs). Cell-free therapy using EVs is an active and emerging field in regenerative medicine. Typical 2D cultures on tissue culture plastic is far removed from the physiological environment of MSCs. The application of 3D cell culture allows MSCs to adapt to their cellular environment which, in turn, influences their paracrine signalling activity. In this study we evaluated the impact of 3D MSCs culture on EVs secretion, cargo proteome composition, and functional assessment in immunomodulatory, anti-inflammatory and anti-fibrotic properties.MSC-EVs from 2D and 3D cultures expressed classical EV markers CD81, CD63, and CD9 with particle diameter of <100 nm. There were distinct changes in immunomodulatory potencies where 3D cultures exhibited reduced indoleamine 2,3-dioxygenase (IDO) activity and significantly reduced macrophage phagocytosis. Administration of 2D and 3D EVs following double dose bleomycin challenge in aged mice showed a marked increase of bodyweight loss in 3D group throughout days 7–28. Histopathological observations of lung tissues in 3D group showed increased collagen deposition, myofibroblast differentiation and leukocytes infiltrations. Assessment of lung mechanics showed 3D group did not improve lung function and instead exhibited increased resistance and tissue damping. Proteome profiling of MSC-EV composition revealed molecular enrichment of EV markers (compared to parental cells) and differential proteome between EVs from 2D and 3D culture condition associated with immune-based and fibrosis/extracellular matrix/membrane organization associated function.This study provides insight into distinct variation in EV protein composition dependent on the cellular microenvironment of the parental cells, which could have implications in their therapeutic effect and potency. Overall, this work suggests that EVs produced from 3D MSC cultures did not enhance typical MSC-EV properties expected from 2D cultures (immunomodulation, anti-fibrotic, anti-inflammatory). The outcome highlights critical differences between MSC-EVs obtained from different culture microenvironments, which should be considered when scaling up MSC culture for clinical manufacturing.

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Section: Introductionmentioning

confidence: 99%

Effect of 2D and 3D Culture Microenvironments on Mesenchymal Stem Cell-Derived Extracellular Vesicles Potencies

Kusuma

Zhu

et al. 2022

Front. Cell Dev. Biol.

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“…In addition, it is indicated that co-culture could induce MSC chondrogenesis potential with increasing Sox9 gene and protein expression, gene expression of ACAN and COL2A1, and improve GAG production 24 . Moreover, studies have demonstrated that aggregation of MSC improves the anti-inflammatory and immunomodulatory effects potential 47 , 48 . These data are consistent with our in vitro and in vivo results.…”

Section: Discussionmentioning

confidence: 99%

Co-aggregation of MSC/chondrocyte in a dynamic 3D culture elevates the therapeutic effect of secreted extracellular vesicles on osteoarthritis in a rat model

Esmaeili

Hosseini

Kamali

et al. 2022

Sci Rep

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Extracellular vesicles (EVs) have therapeutic effects on osteoarthritis (OA). Some recent strategies could elevate EV's therapeutic properties including cell aggregation, co-culture, and 3D culture. It seems that a combination of these strategies could augment EV production and therapeutic potential. The current study aims to evaluate the quantity of EV yield and the therapeutic effect of EVs harvested from rabbit mesenchymal stem cells (MSCs) aggregates, chondrocyte aggregates, and their co-aggregates in a dynamic 3D culture in a rat osteoarthritis model. MSC and chondrocytes were aggregated and co-aggregated by spinner flasks, and their conditioned medium was collected. EVs were isolated by size exclusion chromatography and characterized in terms of size, morphology and surface markers. The chondrogenic potential of the MSC-ag, Cho-ag and Co-ag EVs on MSC micromass differentiation in chondrogenic media were assessed by qRT-PCR, histological and immunohistochemical analysis. 50 μg of MSC-ag-EVs, Cho-ag-EVs and Co-ag-EVs was injected intra-articularly per knee of OA models established by monoiodoacetate in rats. After 8 weeks follow up, the knee joints were harvested and analyzed by radiographic, histological and immunohistochemical features. MSC/chondrocyte co-aggregation in comparison to MSC or chondrocyte aggregation could increase EV yield during dynamic 3D culture by spinner flasks. Although MSC-ag-, Cho-ag- and Co-ag-derived EVs could induce chondrogenesis similar to transforming growth factor-beta during in vitro study, Co-ag-EV could more effectively prevent OA progression than MSC-ag- and Cho-ag-EVs. Our study demonstrated that EVs harvested from the co-aggregation of MSCs and chondrocytes could be considered as a new therapeutic potential for OA treatment.

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“…MSCs are fundamentally responsive to subtle changes in their environment. MSCs respond to changes in atmospheric gases ( Lin et al, 2014 ; Gorgun et al, 2021 ; Roemeling-Van Rhijn et al, 2013 ; Ejtehadifar et al, 2015 ; Kang et al, 2019 ; von Bahr et al, 2019 ), temperature ( Stolzing et al, 2006 ; Kubrova et al, 2020 ; Shimoni et al, 2020 ), hydrostatic pressure ( Steward et al, 2012 ; Becquart et al, 2016 ; Pattappa et al, 2019 ) and aggregation ( Robb et al, 2019 ; Yuan et al, 2019 ; Burand et al, 2020 ; Xie et al, 2021 ). It is surprising then, that the steps and duration between dose preparation and delivery of MSC therapies are ill-defined and under-reported.…”

Section: Discussionmentioning

confidence: 99%

From Vial to Vein: Crucial Gaps in Mesenchymal Stromal Cell Clinical Trial Reporting

Wiese

Wood

Braid

2022

Front. Cell Dev. Biol.

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Retrospective analysis of clinical trial outcomes is a vital exercise to facilitate efficient translation of cellular therapies. These analyses are particularly important for mesenchymal stem/stromal cell (MSC) products. The exquisite responsiveness of MSCs, which makes them attractive candidates for immunotherapies, is a double-edged sword; MSC clinical trials result in inconsistent outcomes that may correlate with underlying patient biology or procedural differences at trial sites. Here we review 45 North American MSC clinical trial results published between 2015 and 2021 to assess whether these reports provide sufficient information for retrospective analysis. Trial reports routinely specify the MSC tissue source, autologous or allogeneic origin and administration route. However, most methodological aspects related to cell preparation and handling immediately prior to administration are under-reported. Clinical trial reports inconsistently provide information about cryopreservation media composition, delivery vehicle, post-thaw time and storage until administration, duration of infusion, and pre-administration viability or potency assessments. In addition, there appears to be significant variability in how cell products are formulated, handled or assessed between trials. The apparent gaps in reporting, combined with high process variability, are not sufficient for retrospective analyses that could potentially identify optimal cell preparation and handling protocols that correlate with successful intra- and inter-trial outcomes. The substantial preclinical data demonstrating that cell handling affects MSC potency highlights the need for more comprehensive clinical trial reporting of MSC conditions from expansion through delivery to support development of globally standardized protocols to efficiently advance MSCs as commercial products.

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Controlled Aggregation Enhances Immunomodulatory Potential of Mesenchymal Stromal Cell Aggregates

Cited by 19 publications

References 89 publications

Effect of 2D and 3D Culture Microenvironments on Mesenchymal Stem Cell-Derived Extracellular Vesicles Potencies

Effect of 2D and 3D Culture Microenvironments on Mesenchymal Stem Cell-Derived Extracellular Vesicles Potencies

Co-aggregation of MSC/chondrocyte in a dynamic 3D culture elevates the therapeutic effect of secreted extracellular vesicles on osteoarthritis in a rat model

From Vial to Vein: Crucial Gaps in Mesenchymal Stromal Cell Clinical Trial Reporting

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