Dose and Time-Dependent Lipopolysaccharide Exposure on A549 Cell Model Influences Pro-Inflammatory Cytokine Interleukin 8

Hyperinflammation in COVID-19 patients is one of the causes of the high mortality rate of COVID-19. An in vitro model mimicking the inflammatory responses in COVID-19 patients is important in the efforts of finding new drug candidates for this disease. Lipopolysaccharide (LPS) can increase the proinflammatory cytokine interleukin 8 in response to the presence of foreign substances. This preliminary study sought to explore the use of  the A549 cells as an in vitro inflammatory model. This study was conducted from August to November 2022 at the stem cell research and development laboratory of Bio Farma Indonesia. The exposure of 100, 500, and 1000 g/mL doses of LPS administered for 24, 72, and 120 hours on the A549 cells was analyzed for cell viability, population doubling time (PDT), and the presence of  proinflammatory cytokine IL-8. The group differences were examined using one- and two-way analysis of variance in IBM SPSS Statistics Version 29, with a p-value of 0.05 considered significant. Cells exposed to a dose of 1000 g/mL LPS had a lower viability and a higher proliferation rate (p<0.05) based on the viability and PDT. Viability, PDT, and pro-inflammatory cytokines showed concentration- and time-dependent responses. Therefore, increased levels of the proinflammatory cytokine IL-8 in cells exposed to LPS at a dose of 1000 g/mL for 24 hours can be used as a mimic to study hyperinflammation in COVID-19 patients

Comparative Preliminary Analysis of Umbilical Cord Blood-, Healthy Adult Peripheral Blood- and Myocardial Infarct Peripheral Blood-Derived Endothelial Progenitor Cells

BACKGROUND: Cardiovascular disease is a leading cause of death globally with the 287,000 deaths per years, characterized by declining of heart function caused by the reduction of heart capacity and lead to heart failure. Cell therapy using endothelial progenitor cells (EPCs) has a big potential for cardiovascular regeneration. EPCs are cells that have ability to differentiate into endothelial cells that can be mobilized and integrated into the defected blood vessel by angiogenesis. AIM: We aimed to seek the superior EPCs derived from MNCs for functional improvement of advanced heart failure patient by cell therapy using EPCs. MATERIALS AND METHODS: We did preliminary analysis to compare umbilical cord blood (UCB), healthy adult peripheral blood (PB)-, and myocardial infarct PB-derived EPCs characteristic and surface phenotypes. Different sources of each EPCs mononuclear cells were characterized by the expression of endothelial (cluster of differentiation [CD] 31, acethylated low-density lipoprotein, and von Willebrand) and hematopoietic stem cell (CD45, CD34, and CD133) surface markers with flow cytometry. RESULTS: In this study, EPCs and the conditioned medium (CM) have been produced and characterized in laboratory scale by comparing several sources of EPCs for instance UCB, PB from healthy people, and patients with myocardial infarction. We have shown that EPC characterizations from each group were not significantly different, but vascular endothelial growth factor and hepatocyte growth factor in UBC-derived CM-EPCs were higher than in PB. CONCLUSION: In conclusion, the UBC-derived EPCs might have a better potential for cardiovascular regeneration

Potential of Conditioned Medium of hATMSCs in Aging Cells Model

Skin aging is caused by the exposure cumulative of ultraviolet radiation, it leads reactive oxygen species (ROS) production in the skin. The conditioned medium of human Adipose Tissue-derived Mesenchymal Stem Cells (hATMSCs) can scavenge free radicals and increase the survival rate of skin cells under oxidative stress. This study examined the protective effects of Conditioned Medium (CM) of hATMSCs in H2O2-induced human skin fibroblast cell line (BJ). The aging cells model using H2O2-induced BJ cells were added CM-hATMSCs in concentrations (0, 10, 30%) and incubated in various time, furthermore BJ cells induced by various H2O2 concentrations (0, 50, 100, 200 µM) incubated for 1 h. The anti-aging potential were measured including viability, ROS and collagen levels in BJ cells which treated CM-hATMSCs. The median inhibitory concentration (IC50) of H2O2 on BJ cells for 1 h incubation was 107.87 μM and 91.25 μM for 10 min incubation. CM-hATMSCs increased the viability on aging model cells. CM-hATMSCs concentration 30% increased the viability of H2O2 50, 100, 200 µM-induced BJ cells. CM-hATMSCs concentration 25% decreased ROS, increased collagen level in H2O2 50, 100, 200 µM-induced BJ cells. CM-hATMSCs increase the viability cells, collagen level and decrease ROS level in aging model cells