The Second Affiliated Hospital of Xi’an Jiaotong University 1 article published in JoVE Biology Isolation and Culture of Primary Cochlear Hair Cells from Neonatal Mice Kun-Yi Wu*1, Bo-Tao Wang*2, Yun-Jia He2, Jing-Yi Xie2, Zi-Chen Chen2, Ying Gao2 1Department of Core Research Laboratory, The Second Affiliated Hospital of Xi’an Jiaotong University, 2Department of Otolaryngology, The Second Affiliated Hospital of Xi’an Jiaotong University Herein, we present a detailed protocol for isolating and culturing primary cochlear hair cells from mice. Initially, the organ of Corti was dissected from neonatal (aged 3-5 days) murine cochleae under a microscope. Subsequently, cells were enzymatically digested into a single-cell suspension and identified using immunofluorescence after several days in culture.