关键词: 人牙周膜细胞；组织块法；供体年龄；细胞培养技术

Abstract:

Abstract: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 cells 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.

Key words: human periodontal ligament cells;tissue method, donor age;cell culture techniques