Human adipose tissue-derived mesenchymal stem cells (hADSCs) due to easy extraction, relative abundance, in vitro expansion and differentiation potential, frozen storage capability, and ability to secrete cytokines, compared to other stem cells, are appropriate candidate in regenerative medicine. Extremely low-frequency electromagnetic fields (ELF-EMF) and betaine are two safe factors in bone lesions repair. This study was designed to assess the osteogenic differentiation potential of these factors on hADSCs. The samples were collected from women undergoing liposuction after obtaining written consent. The hADSCs were extracted and treated with osteogenesis differentiation medium (OD) as the positive control, with OD and betaine (BET group), with OD and EMF (EMF group), and with OD and betaine and EMF (BET+EMF group) for 21 d; the negative control consisted of cells without treatment. Betaine 10 mM and EMF with 50-Hz frequency, 1-mT intensity (8 h daily), and in the form of sinus wave were used. Osteogenic differentiation was evaluated by Alizarin Red staining, alkaline phosphatase activity, calcium deposition, and real-time PCR. A significant increase in calcium deposition in the BET+EMF group was observed compared to the other groups. The activity of alkaline phosphatase in the positive control and BET groups was increased significantly compared to EMF and BET + EMF groups and a significant increase of this enzyme activity in the BET + EMF compared to EMF group was observed. The expression of RUNX2 and OCN genes in the EMF-treated groups were significantly reduced compared to the non-EMF-treated groups, and BET+EMF showed a significant increase of RUNX2 gene expression as compared the EMF group. The ELF-EMF leads to a decrease in the osteogenic differentiation and the expression RUNX2 and OCN genes in hADSCs. But osteogenic differentiation and RUNX2 gene expression were increased post-induction by betaine. The synergic effect of betaine and EMF on the osteogenic differentiation and related genes expression of hADSCs was higher than EMF.

与其他干细胞相比，人类脂肪组织来源的间充质干细胞 (hADSCs) 由于易于提取、相对丰度、体外扩增和分化潜力、冷冻储存能力和分泌细胞因子的能力，是再生医学的合适候选者。极低频电磁场（ELF-EMF）和甜菜碱是骨损伤修复的两个安全因素。本研究旨在评估这些因子对 hADSCs 的成骨分化潜力。样本是在获得书面同意后从接受抽脂的妇女身上收集的。提取hADSCs，以成骨分化培养基（OD）为阳性对照，OD加甜菜碱（BET组），OD加EMF（EMF组），OD加甜菜碱加EMF（BET+EMF组） 21天；阴性对照由未经处理的细胞组成。甜菜碱 10 mM 和 EMF 频率为 50-Hz，强度为 1-mT（每天 8 小时），并以正弦波的形式使用。通过茜素红染色、碱性磷酸酶活性、钙沉积和实时 PCR 评估成骨分化。与其他组相比，观察到 BET+EMF 组的钙沉积显着增加。与 EMF 和 BET + EMF 组相比，阳性对照组和 BET 组中碱性磷酸酶的活性显着增加，并且与 EMF 组相比，观察到 BET + EMF 中的这种酶活性显着增加。的表达 通过茜素红染色、碱性磷酸酶活性、钙沉积和实时 PCR 评估成骨分化。与其他组相比，观察到 BET+EMF 组的钙沉积显着增加。与 EMF 和 BET + EMF 组相比，阳性对照组和 BET 组中碱性磷酸酶的活性显着增加，并且与 EMF 组相比，观察到 BET + EMF 中的这种酶活性显着增加。的表达 通过茜素红染色、碱性磷酸酶活性、钙沉积和实时 PCR 评估成骨分化。与其他组相比，观察到 BET+EMF 组的钙沉积显着增加。与 EMF 和 BET + EMF 组相比，阳性对照组和 BET 组中碱性磷酸酶的活性显着增加，并且与 EMF 组相比，观察到 BET + EMF 中的这种酶活性显着增加。的表达 与 EMF 和 BET + EMF 组相比，阳性对照组和 BET 组中碱性磷酸酶的活性显着增加，并且与 EMF 组相比，观察到 BET + EMF 中的这种酶活性显着增加。的表达 与 EMF 和 BET + EMF 组相比，阳性对照组和 BET 组中碱性磷酸酶的活性显着增加，并且与 EMF 组相比，观察到 BET + EMF 中的这种酶活性显着增加。的表达与非 EMF 处理组相比，EMF 处理组中的RUNX2和OCN基因显着降低，与 EMF 组相比，BET+EMF 显示RUNX2基因表达显着增加。ELF-EMF 导致hADSCs 中成骨分化和RUNX2和OCN基因表达的减少。但甜菜碱诱导后成骨分化和RUNX2基因表达增加。甜菜碱和EMF对hADSCs成骨分化及相关基因表达的协同作用高于EMF。