Effects of insulin on pharmacodynamics of immunosuppressive drugs against mitogen-activated human peripheral blood mononuclear cells

Chen, Shuiling; Sugiyama, Kentaro; Inamura, M; Tanaka, Sachiko; Onda, Kenji; Yin, Hui-jun; Hirano, Toshihiko

doi:10.1080/08923973.2016.1214143

Cited by 5 publications

(3 citation statements)

References 14 publications

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“…In this assay, adapted from Chen et al 2016 [45], PBMCs from 2 different donors, were seeded at 10 6 cells/mL in C-10 medium (RPMI 1640 without phenol red, +10% FBS (Corning, Corning, NY, USA), +1% P/S (Gibco-BRL), +0.1% β-mercaptoethanol (Gibco-BRL, Waltham, MA, USA), +0.1% sodium pyruvate (Gibco-BRL, Waltham, MA, USA), +0.1% sodium bicarbonate (Gibco-BRL, Waltham, MA, USA) and stimulated with 100 pg/mL of LPS. Cells were treated/stimulated in the same way as in the previous assay, with the same test groups and controls but the assay was kept for 96 h. After 96 h, 10 µL/well of an aqueous solution (5 mg/mL) of tetrazolium blue (Merk, Darmstadt, Germany) was added.…”

Section: Mtt Assaymentioning

confidence: 99%

Cytokine Profile and Anti-Inflammatory Activity of a Standardized Conditioned Medium Obtained by Coculture of Monocytes and Mesenchymal Stromal Cells (PRS CK STORM)

et al. 2022

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Intercellular communication between monocytes/macrophages and cells involved in tissue regeneration, such as mesenchymal stromal cells (MSCs) and primary tissue cells, is essential for tissue regeneration and recovery of homeostasis. Typically, in the final phase of the inflammation-resolving process, this intercellular communication drives an anti-inflammatory immunomodulatory response. To obtain a safe and effective treatment to counteract the cytokine storm associated with a disproportionate immune response to severe infections, including that associated with COVID-19, by means of naturally balanced immunomodulation, our group has standardized the production under GMP-like conditions of a secretome by coculture of macrophages and MSCs. To characterize this proteome, we determined the expression of molecules related to cellular immune response and tissue regeneration, as well as its possible toxicity and anti-inflammatory potency. The results show a specific molecular pattern of interaction between the two cell types studied, with an anti-inflammatory and regenerative profile. In addition, the secretome is not toxic by itself on human PBMC or on THP-1 monocytes and prevents lipopolysaccharide (LPS)-induced growth effects on those cell types. Finally, PRS CK STORM prevents LPS-induced TNF-A and IL-1Β secretion from PBMC and from THP-1 cells at the same level as hydrocortisone, demonstrating its anti-inflammatory potency.

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Section: Mtt Assaymentioning

confidence: 99%

Cytokine Profile and Anti-Inflammatory Activity of a Standardized Conditioned Medium Obtained by Coculture of Monocytes and Mesenchymal Stromal Cells (PRS CK STORM)

et al. 2022

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“…First, to test in vitro safety, an MTT-type assay was performed on THP-1 cells transformed to macrophages by adapting the method of Chen et al in 2016 [30]. Differentiated THP-1 cells were washed three times with 0.2 mL of tempered THP-1 medium without PMA and allowed to rest for 30 min before LPS stimuli.…”

Section: Thp-1 In Vitro Inflammation Modelmentioning

confidence: 99%

Evaluation in a Cytokine Storm Model In Vivo of the Safety and Efficacy of Intravenous Administration of PRS CK STORM (Standardized Conditioned Medium Obtained by Coculture of Monocytes and Mesenchymal Stromal Cells)

et al. 2022

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Our research group has been developing a series of biological drugs produced by coculture techniques with M2-polarized macrophages with different primary tissue cells and/or mesenchymal stromal cells (MSC), generally from fat, to produce anti-inflammatory and anti-fibrotic effects, avoiding the overexpression of pro-inflammatory cytokines by the innate immune system at a given time. One of these products is the drug PRS CK STORM, a medium conditioned by allogenic M2-polarized macrophages, from coculture, with those macrophages M2 with MSC from fat, whose composition, in vitro safety, and efficacy we studied. In the present work, we publish the results obtained in terms of safety (pharmacodynamics and pharmacokinetics) and efficacy of the intravenous application of this biological drug in a murine model of cytokine storm associated with severe infectious processes, including those associated with COVID-19. The results demonstrate the safety and high efficacy of PRS CK STORM as an intravenous drug to prevent and treat the cytokine storm associated with infectious processes, including COVID-19.

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“…To test the possible in vitro immunogenicity reaction, an MTTtype assay was performed on macrophage-transformed THP-1 cells by adapting the method of Chen et al [22]. Differentiated THP-1 cells were washed three times with 0.2mL of tempered THP-1 medium without PMA and allowed to stand for 30min before LPS…”

Section: Mtt Assay (3-(34 -Dimethylthiazol-2-thiazolyl)-25diphenyltet...mentioning

confidence: 99%

Combined in Silico and Experimental Research to Test the Possible Immunogenicity of a New Complex Biologic Medical Drug (PRS CK STORM)

Lapuente¹

2022

AJBSR

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Effects of insulin on pharmacodynamics of immunosuppressive drugs against mitogen-activated human peripheral blood mononuclear cells

Cited by 5 publications

References 14 publications

Cytokine Profile and Anti-Inflammatory Activity of a Standardized Conditioned Medium Obtained by Coculture of Monocytes and Mesenchymal Stromal Cells (PRS CK STORM)

Cytokine Profile and Anti-Inflammatory Activity of a Standardized Conditioned Medium Obtained by Coculture of Monocytes and Mesenchymal Stromal Cells (PRS CK STORM)

Evaluation in a Cytokine Storm Model In Vivo of the Safety and Efficacy of Intravenous Administration of PRS CK STORM (Standardized Conditioned Medium Obtained by Coculture of Monocytes and Mesenchymal Stromal Cells)

Combined in Silico and Experimental Research to Test the Possible Immunogenicity of a New Complex Biologic Medical Drug (PRS CK STORM)

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