H. Sun’s scientific contributions

BACKGROUND: There has been no specific method for culturing fibroblasts from cochlear duct membranes so far. OBJECTIVE: To provide good cell model in vitro via culturing fibroblasts from lateral wall of cochlear duct membrane of neonatal C57 mouse by enzyme digestion and differential adhesion methods and to identify its biological features. METHODS: Tissues from lateral wall of cochlear duct membrane were acquired from neonatal C57 mouse using operating microscope. Fibroblasts from lateral wall of cochlear duct membrane were cultured by trypsin digestion combined with differential adhesion methods. The growth condition was observed by inverted phase contrast microscope and hematoxylin-eosin staining, and cell growth curve was drawn, then the immunocytochemistry was employed to classify cell types. RESULTS AND CONCLUSION: After passage and purification, the shape of fibroblasts from lateral wall of cochlear duct membrane was fusiform and triangle, with "S" type cell growth curve. Immunocytochemiscal detection showed that vimentin could be detected in cultured cells, and brown cytoplasmic pigment could be seen. The results demonstrated that fibroblasts can be cultured form lateral wall of cochlear duct membrane of mice.

BACKGROUND: At present, there are differences in reporting culture condition of cochleal spiral ganglion cells (SGCs) in circle of primary cell culture. Individual method has poor repeatability, and is not beneficial for practical application. OBJECTIVE: To primary culture and evaluate SGCs of neonatal C57 mice. METHODS: Tissues from cochlear modiolus were acquired from neonatal C57 mice by microanatomy. SGCs from cochlear modiolus were cultured by trypsin digestion, differential velocity adherent technique combined with chemicals. Cell growth was observed by inverted phase contrast microscope and hematoxylin and eosin staining. Immunocytochemistry was employed to classify cell types. RESULTS AND CONCLUSION: After purification, the cell body of SGCs from cochlear modiolus was ellipse or triangle, with slender processes in cytoplasm. Nuclei were positive for Nuen (stained brown). Cytoplasm and axon were positive for β3-Tubulin (stained brown). These indicated that mouse SGCs were successfully cultured.