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首页> 外文期刊>Experimental and therapeutic medicine >Isolation, culture and induced differentiation of rabbit mesenchymal stem cells into osteoblasts
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Isolation, culture and induced differentiation of rabbit mesenchymal stem cells into osteoblasts

机译：兔间充质干细胞对成骨细胞的分离，培养和诱导分化

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Mesenchymal stem cells (MSCs) may be easily isolated from the bone marrow, and possess multi-lineage differentiation potential and various therapeutic applications. The differentiation of MSCs into osteoblasts is a complex process that is regulated by multiple internal and external factors. In the present study, the differentiation of MSCs isolated from rabbit bone marrow into osteoblasts using different osteoblast inductive media in the presence of dexamethasone, bone morphogenetic protein 2 (BMP-2), 1,25-dihydroxyvitamin D3, transforming growth factor (TGF), platelet lysate and cyclooxygenase 2 (COX2), respectively. Alkaline phosphatase (ALP) activity, mineralization, collagen type (Ct) I and osteocalcin activities, and the mRNA and protein expression levels of vascular endothelial growth factor (VEGF), BMP-2 and Ct II were measured during the differentiation process in MSCs treated with different inducers. Rabbit MSCs were successfully isolated and were observed to be predominantly circular in shape after culture for 24 h. Following subculture for 5 days, the cells demonstrated a spindle shape. ALP, Ct I and osteocalcin activities were higher in cells cultured in dexamethasone, BMP-2 and TGF compared with the activities in control cells. Following differentiation, the dexamethasone, BMP-2 and TGF groups demonstrated significantly enhanced mineralization of MSCs detected by Alizarin Red S staining. The mRNA and protein expression levels of VEGF, BMP-2 and Ct II were significantly increased in the same groups compared with the levels in the control group. In conclusion, rabbit MSCs were successfully isolated from bone marrow and differentiated into osteoblasts indicated by raised ALP, Ct I and osteocalcin activities, mineralization and expression of osteogenesis-inducing genes and proteins. The present study revealed that dexamethasone, BMP-2 and TGF have a positive effect on cell differentiation.
机译：间充质干细胞（MSCs）可以容易地与骨髓分离，并具有多谱分化电位和各种治疗应用。 MSCs进入成骨细胞的分化是由多个内部和外部因素调节的复杂过程。在本研究中，在地塞酮，骨形态发生蛋白2（BMP-2），1,25-二羟基苯胺D3的存在下，使用不同的成骨细胞感应介质将从兔骨髓中分离成骨细胞中的MSC分化到成骨细胞中。转化生长因子（TGF） ，血小板裂解物和环加氧基酶2（COX2）。碱性磷酸酶（ALP）活性，矿化，胶原蛋白（CT）I和骨钙素活动，以及血管内皮生长因子（VEGF），BMP-2和CT II的mRNA和蛋白表达水平在MSC处理的分化过程中测量用不同的诱导者。成功分离兔MSCs，并观察到培养后24小时以培养物的形状主要圆形。亚培养后5天，细胞显示出轴形。与对照细胞中的活性相比，在地塞米松，BMP-2和TGF中培养的细胞中，ALP，CT I和Osteocalcin活性较高。在分化之后，地塞米松，BMP-2和TGF基团显示出通过茜素红S染色检测到的MSCs的显着增强的矿化。与对照组的水平相比，VEGF，BMP-2和CT II的mRNA和蛋白表达水平在相同的基团中显着增加。总之，兔MSCs从骨髓中成功分离，并分化成升高的ALP，CT和骨核苷酸活性，矿化和成骨诱导基因和蛋白质表达的成骨细胞。本研究表明，地塞米松，BMP-2和TGF对细胞分化具有积极影响。

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