人结合上皮和牙龈上皮细胞的培养及生物学特性比较

Culture and comparison of the biological characteristics of human junctional epithelium and gingival epithelium

目的:比较人结合上皮(junctional epithelium,JE)与牙龈上皮(gingival epithelium,GE)的生物学特性。方法:采用细胞培养和免疫组化方法培养鉴定人JE细胞,研究其生物学性质,并与培养的人GE细胞进行比较。结果:人JE细胞形态多样,大小不等,连接疏松,核分裂像多见;而人GE细胞呈典型的角质细胞形态,大小均匀,排列紧密,呈“铺路石”状。免疫组化CK-Pan染色,2种细胞均为阳性,CK19染色JE细胞为阳性,而GE仅见少量散在阳性细胞。生长曲线显示,JE细胞潜伏期(约7d)较GE细胞(约3d)长;进入指数增生期后,JE细胞迅速增殖达高峰(约4d),衰退较快;而GE细胞则匀速增殖达顶点(约7d),平缓下降。JE细胞倍增时间48～60h,可传5代;GE细胞倍增时间72～96h,可传7代。结论:人JE细胞是一种不同于GE细胞的分化程度较低的独特细胞,在本实验培养条件下,人JE细胞的代数少于GE,影响对其深入研究,有待改进培养条件和方法。

PURPOSE： To study the different biological characteristics of human junctional epithelium （JE） and gingival epithelium （GE）.METHODS： Human JE cells were cultured and identified by the cell culture and immunohistochemistry, and the biological characteristics of JE cells and GE cells were compared. RESULTS： The morphology of cultured JE cells was various and unequal, the arrangement of the cells was loose and mitosis was common, while GE colony was consisted of equal and closely packed epithelial-like cells in a paving stone arrangement. CK-Pan staining was positive in all JE and GE cells. CK19 was strongly stained in all JE cells but only moderately stained in some GE cells, and most GE cells were negative which was obviously different from JE cells. JE cells had longer latent period （7 days） than GE cells （4 days） during the cell growth period, and then the cells proliferated rapidly （4 days） to attain the maximum and descended rapidly in the declining period. While GE cells ascended evenly （7 days） to attain the maximum and descended slowly in the declining period. Proliferation study demonstrated the doubling time of JE cells was 48 to 60 hours and that of GE cells was 72 to 96 hours. It was possible to subculture JE cells up to 5 times serially, and that of GE cells was up to 7 times. CONCLUSIONS： The human JE cells are a kind of unique non-differentiated epithelial cells different from GE cells. In this experimental culture condition the subculture times of JE cells were less than GE cells,which affects JE cells, so the culture methods and conditions should be improved.