A streamlined proliferation assay using mixed lymphocytes for evaluation of human mesenchymal stem cell immunomodulation activity

Herzig, Maryanne C.; Delavan, Christopher P.; Jensen, Katherine; Cantu, Carolina; Montgomery, Robbie K.; Christy, Barbara A.; Andrew, P.; Bynum, James A.

doi:10.1016/j.jim.2020.112915

Cited by 17 publications

(20 citation statements)

References 30 publications

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“…To assay responsiveness to mitogenic signaling, PBMCs were plated in triplicate at 300,000 cells/well in a 96-well plate with or without phytohemagglutinin P (PHA-P) at a final concentration of 10 μg/mL. After 72 hours in culture, cells were harvested, and ATP was analyzed by Cell Titer Glo assay as previously described 13 as a measure of cell proliferation.…”

Section: Methodsmentioning

confidence: 99%

High-dose radiation exposure of cold-stored whole blood does not affect hemostatic function

Becerra

Darlington

Abaasah

et al. 2022

J Trauma Acute Care Surg

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Section: Methodsmentioning

confidence: 99%

High-dose radiation exposure of cold-stored whole blood does not affect hemostatic function

Becerra

Darlington

Abaasah

et al. 2022

J Trauma Acute Care Surg

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“…In this study, CM from human primary ADSCs preconditioned with MNPs coated with citric acid and antioxidant protocatechuic acid were tested for their effects on immunomodulatory cytokine release and in limiting cellular responses initiated in allogeneic human PBMC by a known allergen (lectin) using a modified protocol [ 19 ].…”

Section: Discussionmentioning

confidence: 99%

“…For MLR, we used the modified protocol by Herzig et al [ 19 ]. First, 5 mL complete blood was collected in a vacutainer containing citrate as anticoagulant.…”

Section: Methodsmentioning

confidence: 99%

Towards Regenerative Audiology: Immune Modulation of Adipose-Derived Mesenchymal Cells Preconditioned with Citric Acid-Coated Antioxidant-Functionalized Magnetic Nanoparticles

Cumpata

Peptanariu²,

Lungoci³

et al. 2023

Medicina

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Introduction and Background: Based on stem cells, bioactive molecules and supportive structures, regenerative medicine (RM) is promising for its potential impact on field of hearing loss by offering innovative solutions for hair cell rescue. Nanotechnology has recently been regarded as a powerful tool for accelerating the efficiency of RM therapeutic solutions. Adipose-derived mesenchymal cells (ADSCs) have already been tested in clinical trials for their regenerative and immunomodulatory potential in various medical fields; however, the advancement to bedside treatment has proven to be tedious. Innovative solutions are expected to circumvent regulatory and manufacturing issues related to living cell-based therapies. The objectives of the study were to test if human primary ADSCs preconditioned with magnetic nanoparticles coated with citric acid and functionalized with antioxidant protocatechuic acid (MNP-CA-PCA) retain their phenotypic features and if conditioned media elicit immune responses in vitro. MNP-CA-PCA was synthesized and characterized regarding size, colloidal stability as well as antioxidant release profile. Human primary ADSCs preconditioned with MNP-CA-PCA were tested for viability, surface marker expression and mesenchymal lineage differentiation potential. Conditioned media (CM) from ADSCs treated with MNP-CA-PCA were tested for Il-6 and IL-8 cytokine release using ELISA and inhibition of lectin-stimulated peripheral blood monocyte proliferation. Results: MNP-CA-PCA-preconditioned ADSCs display good viability and retain their specific mesenchymal stem cell phenotype. CM from ADSCs conditioned with MNP-CA-PCA do not display increased inflammatory cytokine release and do not induce proliferation of allergen-stimulated allogeneic peripheral blood monocytes in vitro. Conclusions: While further in vitro and in vivo tests are needed to validate these findings, the present results indicated that CM from ADSCs preconditioned with MNP-CA-PCA could be developed as possible cell-free therapies for rescuing auditory hair cells.

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“…In other studies, the functional activity of IDO in MSCs was shown to contribute to inhibiting activated PBMC proliferation due to the depletion of tryptophan and the accumulation of kynurenine and its degradation products [ 35 , 37 , 38 , 39 ]. We observed that, in contrast to COX-2, MSCs did not constitutively express IDO in monoculture.…”

Section: Discussionmentioning

confidence: 99%

Equilibrium among Inflammatory Factors Determines Human MSC-Mediated Immunosuppressive Effect

Suzdaltseva

Goryunov

Silina

et al. 2022

Cells

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Mesenchymal stem cells (MSCs) are thought to be a promising therapeutic agent due to their multiple paracrine and immunomodulatory properties, providing protection from chronic inflammation and promoting tissue repair. MSCs can regulate the balance of pro-inflammatory and anti-inflammatory factors in inflamed tissues, creating a microenvironment necessary for successful healing; however, their interactions with immune cells are still poorly studied. We examined the temporal and spatial changes in gene regulation and the paracrine milieu accompanying the MSC-mediated immunosuppression effect in mixed cultures with activated peripheral blood mononuclear cells (PBMCs). Our data reveal that the peak of suppression of PBMC proliferation was achieved within 48 h following co-culture with MSCs and subsequently did not undergo a significant change. This effect was accompanied by an increase in COX-2 expression and an induction of IDO synthesis in MSCs. At this point, the expression of IL-1, IL-6, IL-8, IFN-γ, MCP-1, and G-CSF was upregulated in co-cultured cells. On the contrary, we observed a decrease in the concentrations of IL-10, IL-13, IL-5, and MIP-1b in co-culture supernatants compared to intact cultures of activated PBMCs. The regulation of IDO, IL-1, IL-6, and G-CSF production was accomplished with the involvement of direct cell–cell contact between MSCs and PBMCs. These findings provide new insights into the use of potential precondition inducers or their combinations to obtain functionally qualified MSCs for more effective treatment of inflammatory diseases.

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A streamlined proliferation assay using mixed lymphocytes for evaluation of human mesenchymal stem cell immunomodulation activity

Cited by 17 publications

References 30 publications

High-dose radiation exposure of cold-stored whole blood does not affect hemostatic function

High-dose radiation exposure of cold-stored whole blood does not affect hemostatic function

Towards Regenerative Audiology: Immune Modulation of Adipose-Derived Mesenchymal Cells Preconditioned with Citric Acid-Coated Antioxidant-Functionalized Magnetic Nanoparticles

Equilibrium among Inflammatory Factors Determines Human MSC-Mediated Immunosuppressive Effect

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