摘要
目的观察正常人关节软骨细胞的体外培养及冻存后再复苏对人体关节软骨细胞活性所产生的影响。方法在无菌条件下取因创伤所致离断且无再植条件残肢的关节软骨组织,采用Ⅱ型胶原酶一次消化的方法获取原代软骨细胞后进行体外培养,并经10%DMSO冻存液冻存及常规复苏,过程中均用倒置显微镜定期观察软骨细胞的形态生长变化以及复苏后细胞的存活率。结果常规消化传代后的第2～4代间贴壁时间无明显差别,但较原代明显变短,生长速度加快,在形态上仍以三角或多角形为主,传代在5代以内的软骨细胞仍能保持正常的组织学形态,且复苏后软骨细胞的存活率能达95%。结论原代培养获得的正常人体关节软骨细胞在5代以内者可以较好地保留软骨细胞的特性和活性。
Objective To accomplish the primary culture of normal articular chondrocyte by one enzyme digestion method and confirm the influence of cryopreservation on the cell activity of human chondrocytes.Methods Harvest the chondrocytes from the articular cartilage of amputated limb due to trauma which can＇t be replanted with collagenase Ⅱ digestion method under sterile conditions.The inverted microscope was used to observe the changes in cell morphology and survival of these chondrocytes after cryopreservation with 10% of DMSO and recovery.Results Two to four intergenerational passage of adherent time is with no significantly difference in conventional digestion,but shorter than the primary,and growth rate accelerates,mainly in triangular or polygonal morphology.The chondrocytes passaged less than five generations can maintain normal morphology and their survival rate can reach 95% after cryopreservation and recovery.Conclusion Cultures of normal human articular chondrocytes in less than five generations can still retain the characteristics and activity.
出处
《中华全科医学》
2012年第7期1001-1003,F0003,共4页
Chinese Journal of General Practice
基金
浙江省台州市科技局项目(08KY15)
关键词
软骨细胞
培养
冻存
复苏
Chondrocyte
Culture
Cryopreservation
Recovery
