Macrophages and fibroblasts are two types of important cells in wound healing. The development of novel platforms for studying the interrelationship between these two cells is crucial for the exploration of wound-healing mechanisms and drug development. In this study, a microfluidic chip composed of two layers was designed for the co-culturing of these two cells. An air valve was employed to isolate fibroblasts to simulate the wound-healing microenvironment. The confluence rate of fibroblasts in the co-culture system with different macrophages was explored to reflect the role of different macrophages in wound healing. It was demonstrated that M2-type macrophages could promote the activation and migration of fibroblasts and it can be inferred that they could promote the wound-healing process. The proposed microfluidic co-culture system was designed for non-contact cell–cell interactions, which has potential significance for the study of cell–cell interactions in biological processes such as wound healing, tumor microenvironment, and embryonic development.
Three-dimension (3D) cell co-cultural spheroids exhibit enhanced cellular functions and they can mirror in-vivo microenvironments. Herein, a sessile drop chip was developed to construct 3D spheroids for mirroring the wound healing microenvironment. The sessile drop chip holds the superhydrophobic surface of each microwell which can facilitate cell suspensions transfer to spheroids through the offset of surface tension and gravity, and each microwell has a cylinder hole that offers adequate oxygen to spheroids. It was demonstrated that the 3T3 fibroblast spheroid and the 3T3 fibroblast/M2-type macrophage co-culture spheroid can be formed and remained the physiological activity within nine days. 3D morphology of spheroids was reconstructed using the transparent processing technology and Z-stack function of confocal microscopy. Characteristics of proliferation and differentiation were analyzed by using nano antibody-based 3D immunostaining assay. Results revealed that M2-type macrophages can promote the proliferation and differentiation of the 3T3 fibroblast spheroid. This study presented a novel affordable platform for developing 3D spheroids and provides a 3D model for investigating the macrophages-associated wound healing microenvironment.
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