论文部分内容阅读
目的:观察骨康方含药血清对三维培养成骨细胞系统生长繁殖、矿化能力和ER-βmRNA表达的变化,探讨成骨细胞三维培养体系在体外中药干预研究中的应用。方法:体外分离成骨细胞,进行二维培养和三维培养,分别用不同浓度阴性血清培养基和药物血清培养基进行培养,使用MTT法检测成骨细胞的生长情况,茜素红染色法观察成骨细胞矿化能力,RT-PCR法检测ER-βmRNA表达的变化。结果:与二维培养体系比较,骨康含药血清对三维体系下培养的成骨细胞的促增殖作用更加明显;细胞ER-beta的mRNA转录水平显著提高;矿化能力更强。结论:三维培养系统比二维培养系统更有利于细胞功能的保持和发挥,更适合于体外中药干预研究中的作用。
OBJECTIVE: To observe the effects of Gukangfang containing serum on the growth, reproduction, mineralization and ER-βmRNA expression of osteoblast in three-dimensional culture system and to explore the application of osteoblast three-dimensional culture system in vitro. Methods: Osteoblasts were isolated in vitro and cultured in two-dimensional and three-dimensional culture. The cells were cultured with different concentrations of serum-free medium and serum-containing medium. The growth of osteoblasts was detected by MTT assay. Alizarin red staining Osteoblast mineralization ability, RT-PCR detection of ER-βmRNA expression changes. Results: Compared with the two-dimensional culture system, Gukang medicated serum had a more obvious effect on promoting the proliferation of osteoblasts cultured in three-dimensional system. The transcription level of ER-beta mRNA was significantly increased and the mineralization ability was stronger. Conclusion: The three-dimensional culture system is more conducive to maintaining and exerting cellular functions than the two-dimensional culture system, which is more suitable for the role of traditional Chinese medicine in vitro intervention research.