Chen L S, Couwenhoven R I, Hsu D, Luo W, Snead M L
Center for Craniofacial Molecular Biology, University of Southern California, School of Dentistry, Los Angeles 90033.
Arch Oral Biol. 1992 Oct;37(10):771-8. doi: 10.1016/0003-9969(92)90110-t.
Electroporation was used to introduce foreign genes into cells derived from the mouse enamel organ epithelia (EOE). Optimal conditions for this electroporation were established. The introduction of a plasmid construct bearing the coding region for the large T-antigen from polyoma virus into EOE cells permitted the establishment of a derivative cell line that has the following characteristics: (1) the cells could be passaged many times; (2) they expressed a keratin-containing cytoskeleton; and (3) approx. 60% of the cells expressed amelogenin, a tissue-specific gene product unique to ameloblasts. Potential uses for such a cell line include analysis of: (1) the upstream regulatory regions required for temporally and spatially restricted expression of amelogenin; (2) the post-translational modification of amelogenin in synchronized cells and (3) the organization and biomineralization of enamel extracellular matrix in monolayer culture.
采用电穿孔法将外源基因导入源自小鼠釉质器官上皮（EOE）的细胞中。确定了该电穿孔的最佳条件。将携带多瘤病毒大T抗原编码区的质粒构建体导入EOE细胞，从而建立了具有以下特征的衍生细胞系：（1）细胞能够传代多次；（2）它们表达含角蛋白的细胞骨架；（3）约60%的细胞表达釉原蛋白，这是成釉细胞特有的一种组织特异性基因产物。这种细胞系的潜在用途包括分析：（1）釉原蛋白在时间和空间上受限表达所需的上游调控区域；（2）同步化细胞中釉原蛋白的翻译后修饰；（3）单层培养中釉质细胞外基质的组织和生物矿化。
