摘要
目的建立兔髓核内细胞的体外培养方法,绘制其生长曲线。方法取一月龄新西兰兔的髓核,胰酶和胶原酶消化,DMEM/F12培养基中培养,倒置显微镜观察细胞形态。MTT法测定细胞OD值以绘制其生长曲线。结果原代培养髓核内细胞多为多角形,另有细胞为圆形或椭圆形,内含空泡。随传代培养的次数增加,细胞形态渐变为梭形,出现反分化的特性。生长曲线显示,髓核内细胞在19d时达到高峰。结论成功建立了兔髓核细胞体外培养体系,为组织工程化椎间盘的研究打下基础。在实际应用时,以采取次代细胞为佳,并应将其培养至第19天。
Objective To establish a method for the culture of rabbit nu cleus pulposus-derived cells and determinate its growth curve. Methods Nucleus pulposus tissue taken from a one-month-old rabbit was treated by Trypsin and collagenase, and then the cells were cultured in DMEM/F12 medium. Cell morpholog y was observed by an invert microscope and cells growth curve was drawn. Results Primary culture cells of the nucleus pulposus were composed of small polygonal cells and large round or ellipse cells with vacuoles. The cells changed morpholo gically into the spindle-shape following the increase of subcultures. In other words, the cells presented the sign of dedifferentiation. At 19th day, the growt h curve showed that the cell proliferation rose up to the top. Conclusions The successful establishment of a method for the culture of rabbit nucleus pulposus-derived cells provides a tissue engineering basis for the study on the interve rtebral disc. In authors’ opinion, subculture cells will be the best choice in the future research.
出处
《中华创伤骨科杂志》
CAS
CSCD
2004年第2期184-186,共3页
Chinese Journal of Orthopaedic Trauma
基金
广东省科委重点攻关项目(2KM055038)
广州市科技攻关项目[(JB02)2K201701]
