Abstract:
The osteogenic differentiation of mesenchymal stem cells is now a standard procedure in modern bone tissue engineering. As this is a promising field for future clinical applications, many cell culture media exist to promote osteogenic differentiation. Prior to differentiation, cells must be expanded to obtain sufficient numbers for experiments. Little evidence is available regarding the optimal media combination for expansion and differentiation to maximize the osteogenic response. Therefore, human BM-MSCs (n = 6) were expanded in parallel in DMEM (Dulbecco\'s Modified Eagle Medium) LG (Low Glucose) and α-MEM (Minimum Essential Media alpha-modification), followed by simultaneous monolayer differentiation toward the osteogenic lineage in: 1. DMEM LG (Low Glucose), 2. DMEM HG (High Glucose), 3. α-MEM, 4. \"Bernese medium\", and 5. \"Verfaillie medium\", with a corresponding negative control (total 20 groups). As a marker for osteogenic differentiation, hydroxyapatite was accessed using radioactive 99mTc-HDP labeling and quantitative alizarin red staining. The results indicate that all media except \"Bernese medium\" are suitable for osteogenic differentiation, while there was evidence that DMEM LG is partly superior when used for expansion and differentiation of BM-hMSCs. Using \"Verfaillie medium\" after DMEM LG expansion led to the highest grade of osteogenic differentiation. Nevertheless, the difference was not significant. Therefore, we recommend using DMEM LG for robust osteogenic differentiation, as it is highly suitable for that purpose, economical compared to other media, and requires little preparation time.
摘要:
间充质干细胞的成骨分化现在是现代骨组织工程中的标准程序。由于这是未来临床应用的一个有希望的领域,存在许多细胞培养基以促进成骨分化。在分化之前,细胞必须扩大,以获得足够的数量进行实验。关于扩增和分化的最佳培养基组合以最大化成骨反应的证据很少。因此,人BM-MSCs(n=6)在DMEM(Dulbecco's改良的Eagle培养基)LG(低葡萄糖)和α-MEM(最低必需培养基α-修饰)中平行扩增,随后同时向成骨谱系进行单层分化:1.DMEMLG(低葡萄糖),2.DMEMHG(高葡萄糖),3.α-MEM,4.“伯尔尼媒体”,和5.\"Verfailliemedium\",与相应的阴性对照(共20组)。作为成骨分化的标志,使用放射性99mTc-HDP标记和定量茜素红染色访问羟基磷灰石。结果表明,除“Bernese培养基”外,所有培养基均适用于成骨分化,而有证据表明,当用于BM-hMSCs的扩增和分化时,DMEMLG部分优越。在DMEMLG扩增后使用“Verfaillie培养基”导致最高等级的成骨分化。然而,差异不显著。因此,我们建议使用DMEMLG进行强大的成骨分化,因为它非常适合这个目的,与其他媒体相比经济,需要很少的准备时间。