Study on the culture and identification of annulus fibrosus cells in rabbit

Objective: To establish and identify the culture model of annulus fibrosus(AF) cells of rabbit in vitro for providing the theoretical basis in further studying the intervertebral disc degeneration.Methods: The primary AF cells were obtained using enzyme digestion method,and the AF cells after passage were purified using gradient centrifugation technique and natural digestion method.The cytological characteristics of AF cells were observed,the growth curve of cells was drew by MTT assay.The expression levels of type Ⅰ and Ⅱ collagen in AF cells were measured using immunofluorescence combined with Hoechst nuclear staining,and the AF cells were identified.Results: The firstly adherent AF cells was round,and soon extended for the short spindle.After passage,the cells was long spindle or polygon,and began to age after fourth generation,and the expressions of type Ⅰand Ⅱ collagen in AF cells were found.Conclusions: The AF cells of rabbit are successfully cultured in vitro,which can provide the basis in further studying the occurrence and development mechanism of intervertebral disc degeneration.