不同原代培养方法培养人牙髓细胞的研究.pdf

2011 39 2
天津医药年 2 月第卷第期 139
不同原代培养方法培养人牙髓细胞的研究
沈晴昳
45
摘要目的:建立一种操作简单方便的人牙髓细胞原代培养方法。方法:选择因正畸拔除的完整无龋双尖牙
15
颗,无菌条件下取出牙髓,分别采用组织块法、酶消化法和改良组织块法(每法颗)进行人牙髓细胞的体外原代培
3 3
养,比较观察种不同培养方法的成活率、细胞形态和数量,测定生长曲线并进行细胞来源鉴定。结果: 种方法均可
从人牙髓组织分离培养获得外胚间充质来源的牙髓细胞,且呈成纤维细胞样,抗波形丝蛋白染色阳性,抗角蛋白染色
4 % , ,2
阴性。其中,组织块法耗时较长, 例培养成功( );酶消化法培养成活率较低且细胞数量较少例培养成功
% 12 %
( );改良组织块法中未发现有漂浮的组织块, 例培养成功( )。结论:改良组织块法解决了传统组织块法
,
中组织块难以贴壁的问题,简化了操作步骤并具有较高的成功率是一种科学可行的人牙髓细胞原代培养方法。
关键词牙髓细胞双尖牙细胞培养技术体外研究
Study on Different Methods in Primary Cultures of Human Dental Pulp Cells in Vitro
SHEN Qingyi
Department of Prosthodontics, Shanghai Stomatological Disease Center, Shanghai 200031, China
To establish a simple primary culture technique of human dental pulp cells in vitro.
Abstract Objective: Methods:
The pulp tissue was separated from 45 healthy young human teeth extracted for orthodontic purpose. The pulp explants were
cultured by three methods, tissue explants method, tissue-explants collagenase/dispase digestion method and modified tissue
culture techniques respectively. The cell culture efficiency cell morphology and quantity were observed and evaluated by
,
phase-contract microscopy and growth curres of human pulptissue cells. Immunocytochemical staining method was used to
characterize the dental pulp cell lineage. The cultured cells appeared to be fibroblast-like