Neutrophils Culture in Collagen Gel System

Abstract

Neutrophils (Neu) migrate rapidly to damaged tissue and play critical roles in host defense and tissue homeostasis, including the intestinal epithelia injuries and immune responses. Although their important roles in these diseases, they are challenging to study due to their short life span and the inability to cryopreserve or expand them in vitro. Moreover, the standard cell culturing on plastic plates (two-dimensional (2D) cultures) does not represent the actual microenvironment where cells reside in tissues. In this study, we developed a new three-dimensional (3D) culture system for human and mouse peripheral blood Neu, which is made of hydrogel. The Neu showed much better cell integrity and less cell debris in the 3D culture system compared to that in 2D culture system. Moreover, the 3D culture system was more suitable for the observation of neutrophil extracellular traps (NETs) stimulated by the classical stimulation phorbol ester (PMA), and other damage associated molecular patterns (DAMPs) such as Lipopolysaccharide (LPS)/ATP, interleukin-1 β (IL-1β) and tumor necrosis factor α (TNFα) than the 2D culture system. Moreover, NETs phenomenon in 3D culture system is similar to that in vivo. In addition, the 3D culture system was evaluated to co-culturing Neu and other parenchymal cells, such as colon mucosal epithelial cell lines. In conclusion, the 3D culture system could maintain better properties of Neu than that in 2D culture system and it may reduce the gap between in vitro an in vivo experimentations.

Heat acclimation with probiotics-based ORS supplementation alleviates heat stroke-induced multiple organ dysfunction via improving intestinal thermotolerance and modulating gut microbiota in rats

“…However, the relationship between the heat shock response, intestinal barrier function, and gut microbiota nevertheless merits further investigation—particularly in terms of HA and HS. It is also worth to note that both HA and HS are involved in the problem of rhabdomyolysis, and whether ORSP supplementation can help solve rhabdomyolysis is also worth verifying in follow-up studies ( He et al, 2020 , 2022 ; Li et al, 2022 ). It has been reported that a kind of probiotic Akkermansia muciniphila could improve heat stress-impaired intestinal barrier function by modulating HSP27, which suggests that it is important to study the relationship between probiotics and HSPs.…”

Biomimetic cell culture for cell adhesive propagation for tissue engineering strategies

“…5 The three main components of the ECM are fibrin, glycoproteins and proteoglycans, which regulate the biochemical and mechanical properties, provide sufficient cell-ECM interactions and mediate cell-cell interactions. 6,7 Therefore, the key to achieving biomimetic cell culture is to recapitulate the ECM in vitro to provide a simulated growth environment for cells as in vivo. 8 In general, the cellular microenvironment in vivo can be approximately divided into three aspects, i.e., the spatial microenvironment, chemical microenvironment, and physical microenvironment (Fig.…”

Data_Sheet_1.ZIP

“…However, the relationship between the heat shock response, intestinal barrier function, and gut microbiota nevertheless merits further investigation-particularly in terms of HA and HS. It is also worth to note that both HA and HS are involved in the problem of rhabdomyolysis, and whether ORSP supplementation can help solve rhabdomyolysis is also worth verifying in follow-up studies (He et al, 2020(He et al, , 2022Li et al, 2022). It has been reported that a kind of probiotic Akkermansia muciniphila could improve heat stressimpaired intestinal barrier function by modulating HSP27, which suggests that it is important to study the relationship between probiotics and HSPs.…”