The Explorating Reform of Cell Biology Experiment Teaching—Extraction of Murine Peritoneal Macrophages and Detection of NF-κB Signaling Pathway Activation

Cell Biology Experiment is an important part of Cell Biology courses and plays an important role in cultivating students’ practice and scientific research thinking abilities. “Cell Signal Transduction” is the key and difficult chapter of Cell Biology course. The detection of intracellular signal transduction is an crucial experiment to the universities’s undergraduate students. Due to the complicated steps of signal transduction experiments and students are not familiar with the procedures, the experimental results are often not satisfactory, and it is difficult for students to observe significant phenomena. Therefore, in the current teaching of undergraduate Cell Biology Experiments in universities, there are few experiments on the detection and observation of intracellular signal transition. The purpose of this study is to investigate the activation of NF-κB signaling pathway in murine peritoneal macrophages. Four interrelated experiments were designed, which were: (1) extraction and purification of murine peritoneal macrophages; (2) observation of the localization of RelA/p65 in cells stimulated by inflammatory factors; (3) the mRNA expression of inflammatory factors in murine peritoneal macrophages were detected; (4) the protein expression of inflammatory factors in murine peritoneal macrophages were detected. This experiment is easy to isolate the murine peritoneal macrophages and observe the obvious relocation of RelA/p65. The independent and complete comprehensive experiment can enrich and optimize the content and model of undergraduate teaching of Cell Biology Experiment. Furthermore, the experiment can help students to truly understand the cell signaling pathways in the inflammatory response process. 

CSTR: 32200.14.cjcb.2024.06.0012