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Fig. 1 | Stem Cell Research & Therapy

Fig. 1

From: Direct comparison of different therapeutic cell types susceptibility to inflammatory cytokines associated with COVID-19 acute lung injury

Fig. 1

GMP stem cell manufacturing and characterization. A Conceptual figure showing COVID-19 ARDS patient lung microenvironment and methodological approach utilized in the study: All three cell products (BM-MSCs, HDCs, & UC-MSCs) were prepared to the clinical-grade standards in the cell manufacturing facility. Using COVID-19 clinical literature survey and flow cytometry cytokine receptor expression profile, five COVID-19 ARDS related cytokines (IL-1β, IL-2, IL-8, IL-10, and TNF-α) were selected for experimentation in the current study. Followed by an assessment of individual cytokine dose response on cell viability, the impact of cytokine cocktail containing all five cytokines at clinically relevant doses on paracrine output (extracellular vesicle production) and cell viability of three cell types was evaluated. All experiments were conducted at hypoxia (1% oxygen) for 48-h to simulate transplanted cell residence in hypoxic COVID-19 ARDS lung microenvironment. B Flow cytometry analysis showed BM-MSCs & UC-MSCs were positive for markers of mesenchymal stromal cell origin (CD73, CD90, CD105, CD166, and CD44) and negative for markers of hematopoietic cell origin (CD14, CD19, CD34, CD45 and HLA-DR). C qPCR showed IFN-γ treatment for 24-h induces IDO-gene expression in all three cell types (GAPDH was used as a reference gene). D Colorimetric assay showed Tissue factor activity of all three cell types was found to be similar. E qPCR showed none of the cell types express ACE2 (ΔCq values < -20 when normalized to reference gene, GAPDH). All data are presented as individual and mean values ± SEM, n = 3 biological replicates; each circle represents one data point from one unique biological replicate

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