3D co-culture of macrophages and fibroblasts in a sessile drop array for unveiling the role of macrophages in skin wound-healing

作者全名："Lyu, Xiaoyan; Cui, Feiyun; Zhou, Hang; Cao, Bo; Zhang, Xiaolan; Cai, Minghui; Yang, Shulong; Sun, Bangyong; Li, Gang"

作者地址："[Lyu, Xiaoyan] Sichuan Univ, West China Hosp, Clin Inst Inflammat & Immunol, Frontiers Sci Ctr Diseaserelated Mol Network,Dept, Chengdu 610041, Peoples R China; [Cui, Feiyun; Cao, Bo; Zhang, Xiaolan; Cai, Minghui] Harbin Med Univ, Sch Basic Med Sci, Harbin 150081, Peoples R China; [Zhou, Hang] Chongqing Med Univ, Sch Lab Med, Key Lab Clin Diagnost, Minist Educ, Chongqing 400016, Peoples R China; [Yang, Shulong] Harbin Med Univ, Affiliated Hosp 6, Dept Pediat Surg, Harbin 150081, Peoples R China; [Sun, Bangyong; Li, Gang] Chongqing Univ, Key Lab Optoelect Technol & Syst, Def Key Disciplines Lab Novel Micronano Devices &, Minist Educ, Chongqing 400044, Peoples R China"

通信作者："Lyu, X (通讯作者)，Sichuan Univ, West China Hosp, Clin Inst Inflammat & Immunol, Frontiers Sci Ctr Diseaserelated Mol Network,Dept, Chengdu 610041, Peoples R China.; Li, G (通讯作者)，Chongqing Univ, Key Lab Optoelect Technol & Syst, Def Key Disciplines Lab Novel Micronano Devices &, Minist Educ, Chongqing 400044, Peoples R China."

来源：BIOSENSORS & BIOELECTRONICS

ESI学科分类：CHEMISTRY

WOS号：WOS:000927081600001

JCR分区：Q1

影响因子：10.7

年份：2023

卷号：225

期号：　

开始页：　

结束页：　

文献类型：Article

关键词：Sessile drop array; Three-dimensional cell co -culture; Wound healing microenvironment; Macrophage modulated fibroblasts

摘要："Three-dimensional (3D) heterotypic multicellular spheroid models play important roles in researches of the proliferation and remodeling phases in wound healing. This study aimed to develop a sessile drop array to cultivate 3D spheroids and simulate wound healing stage in vitro using NIH-3T3 fibroblasts and M2-type mac-rophages. By the aid of the offset of surface tension and gravity, the sessile drop array is able to transfer cell suspensions to spheroids via the superhydrophobic surface of each microwell. Meanwhile, each microwell has a cylinder hole at its bottom that provides adequate oxygen to the spheroid. It demonstrated that the NIH-3T3 fibroblast spheroid and the 3T3 fibroblast/M2-type macrophage heterotypic multicellular spheroid can form and maintain physiological activities within nine days. In order to further investigate the structure without destroying the entire spheroid, we reconstructed its 3D morphology using transparent processing technology and the Z-stack function of confocal microscopy. Additionally, a nano antibody-based 3D immunostaining assay was used to analyze the proliferation and differentiation characteristics of these cells. It found that M2-type mac-rophages were capable of promoting the differentiation of 3T3 fibroblast spheroid. In this study, a novel, inexpensive platform was constructed for developing spheroids, as well as a 3D immunofluorescence method for investigating the macrophage-associated wound healing microenvironment."

基金机构："National Natural Science Foundation of China [81871567, 61974012]"

基金资助正文：This work was supported by the National Natural Science Foundation of China (No. 81871567 and No. 61974012) .