摘要
目的:通过研究供体年龄对组织块法原代培养人牙周膜细胞的影响,寻找人牙周膜细胞原代培养的最佳供体年龄,为便捷、快速地获得大量人牙周膜细胞提供理论依据。方法:将在口腔外科门诊收集正畸拔除的12～28岁青少年健康前磨牙68颗,按供体年龄分为3组:12～14岁组、15～18岁组和19～28岁组,采用组织块法原代培养人牙周膜细胞并传代,通过形态学和免疫化学染色方法对其进行鉴定。结果:不同年龄组牙周膜经组织块法原代培养获得的人牙周膜细胞的成功率不同,12～14岁组最高,成功率为66.67%;15～18岁组次之,成功率为8.57%;19～28岁组最低,成功率为0。12～14岁和15～18岁组传至第3代细胞免疫组织化学检测显示细胞波形丝蛋白染色阳性,角蛋白染色阴性,表明为中胚层组织来源。结论:供体年龄对体外原代培养人牙周膜细胞成功率有很大影响,12～14岁为最佳供体年龄。
Objective Through studying the impact of donor age on the human periodontal ligament cells （PDLC） of primary culture, to find the best donor age of the human PDLC of primary culture, and provid a theoretical basis for convenient, quick acquisition of a large number of human PDLC. Methods 68 adolescent （12--28 years old） health premolar were collected, which were extracted for the reason of orthodontic in Department of Oral and Maxillofacial Surgery. Then the teeth were divided into three groups according to donor age, namely 12--14 years old group, 15--18 years old group and 19- 28 years old group. The primary eells were isolated from human periodontal ligament and cultivated by tissue explant culture technique, then passaged when the cells completely covered the bottom of culture flasks. Morphological analysis and immunocytochemical analysis were used to identify the cell lineage. Results Human PDLC of primary culture were obtained with tissue cultivation, the success rates in various groups were different, The success rates in 12--14 years old group, 15--18 years old group and 19-- 28 years old group were 66.67%, 8.57%, and 0, respectively. The immunohistochemical result showed that when the cells were cultivated to passage 3 in 12--14 years old and 15--18 years old groups, they were positive for vimentin staining, and negative for keratin staining, and they were proved to be tissue derived mesoderm. Conclusion Donor age has a significant effect on the success rate of human PDLC and 12-14 years old is the best donor age.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2010年第1期95-98,I0002,共5页
Journal of Jilin University：Medicine Edition
基金
吉林省科技厅科技发展计划项目资助课题(200705342)
吉林省长春市科技计划项目资助课题(2008107)
关键词
人牙周膜细胞
组织块法
供体年龄
细胞培养技术
human periodontal ligament cells
tissue method
donor age
cell culture techniques
