Int J Stem Cells.  2020 Jul;13(2):212-220. 10.15283/ijsc19155.

Human Amniotic Epithelial Cells Affect the Functions of Neutrophils

• Alipour R
• Motedayyen H
• Sereshki N
• Rafiee M
• Alsahebfosul F
• Pourazar A

• KMID: 2504334
• DOI: http://doi.org/10.15283/ijsc19155

Abstract

Background and Objectives
As a stem cell group, Human amniotic epithelial cells (HAECs) have numerous advantages over their embryonic and adult counterparts for therapeutic utility. They are closer to clinical applications compared to other stem cell types. Additionally, the anti-inflammatory and immunoregulatory properties of HAECs toward several immune cells have been shown previously. Nevertheless, despite the ever-increasing importance of neutrophils in the immune and non-immune processes, a few studies investigated the interaction of neutrophils and HAECs. To increase the current knowledge of HAECs immunology which is necessary for optimizing their future clinical applications, here we explored the effect of HAECs on two chief neutrophil functions; respiratory burst and phagocytosis.
Methods and Results
Freshly isolated human blood neutrophils were co-cultured with different number of HAECs for about 24 or 48 hours, then the oxidative burst and phagocytosis of stimulated neutrophils were assessed and compared. The results demonstrated a substantial elevation in the phagocytosis percentage, conversely a significant reduction in the oxidative burst of HAECs-cocultured neutrophils. These effects were dose-dependent, but did not show similar patterns. Likewise, the elongation of coculture period inversely influenced the HAECs-induced effects on the two neutrophil functions.
Conclusions
The present study, for the first time, investigated the HAECs-mediated effects on the two main neutrophil functions. The findings suggest that HAECs by enhancement of phagocytic ability and simultaneously, attenuation of oxidative burst capacity of neutrophils protect the fetus from both microbial treats and oxidative stress and their consequent inflammation; thus corroborate the current anti-inflammatory vision of HAECs.

Keyword

Human amniotic epithelial cells; Neutrophils; Oxidative burst; Phagocytosis

Figure

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  Fig. 1 HAECs influenced the phagocytosis of human blood granulocytes in cocultures. Isolated blood granulocytes were cocultured with hAECs at various ratio and time (as detailed in the text). After coculture, neutrophils and HAECs were separated by adherence. For phagocytosis assay, the neutrophils were stimulated and added by fluorescent 1.0 μm diameter beads and run on the flow cytometer. The uptake of fluorescent beads causes the neutrophils become fluorescent too; thus the percent of fluorescent granulocytes presented as phagocytosis percentage (A); unstimulated neutrophils were used as negative control (B).

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  Fig. 2 The effects of HAECs number on the functions of granulocytes. The oxidative burst capacity of human granulocytes changed when they were cocultured with different number of HAECs. The levels of respiratory burst were assessed by flow cytometry and for comparison of the results, OBI (oxidative burst index) was calculated; that is the ratio of mean fluorescence of stimulated neutrophils to mean fluorescence of unstimulated neutrophils (A). The coculture of human granulocytes with various number of hAECs altered the phagocytosis percentage of the neutrophils (B).

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  Fig. 3 The impact of HAECs on the oxidative burst and phagocytosis of granulocytes. On the whole, the data analysis revealed that co-cultivation of neutrophils with HAECs reduced the level of oxidative burst (p=0.000) (A); but enhance the phagocytosis (p=0.000) in comparison to neutrophils cultured alone (without hAECs) (one way ANOVA, n=5) (B); hAECs: human amniotic epithelial cells, control (−): resting (unstimulated) neutrophils.

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  Fig. 4 HAECs affected neutrophil functions time- and dose-dependently. The HAECs-induced inhibition of respiratory burst of neutrophils differed in the presence of different number of HAECs and diminished by time. However, the ratio groups do not differ statistically significant in their degree of change in the OBI over time (Repeated measurement ANOVA, n=5, p=0.104) (A). The phagocytic activity of neutrophils were raised considerably in all ratios of HAECs to neutrophils, and the lowest ratio caused the highest level; it also showed an increase by time. Additionally, there was an interaction between time and cell ratio; the effect of cell numbers on the phagocytosis varied over time (Repeated measurement ANOVA, n=5, p=0.034) (B).

Reference

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