UNASSIGNED: Differentiation capacities and stemness potential of harvested ADSCs were evaluated by differentiating into osteocytes and adipocytes, and expression of mesenchymal CD markers by flow cytometry. The optimum dose of crocin was assessed with an MTT assay. Crocin, retinoic acid, CREB/BDNF, and Notch inhibitors and their combination were added to the culture medium. Jag1, Hes1, Notch, and BDNF gene expression were analyzed by RT-PCR on days 7, 14, and 21, while CREB, DCX, SOX2, and NeuN expression were analyzed by immunofluorescence.
UNASSIGNED: Expression of mesenchymal CD markers as well as adipogenic and osteogenic differentiation confirmed the origin and properties of ADSCs. The optimal dose of crocin was 1 mM. Crocin significantly (P<0.05) increased, while inhibitors (DATP&Naphthol) significantly (P<0.05) decreased Jag1, Hes1, Notch, and BDNF expression. Immunofluorescent assessments showed that expression of DCX, BDNF, NeuN, and Sox2 proteins increased significantly (P<0.05) after crocin administration and decreased significantly (P<0.05) after inhibitor administration.
UNASSIGNED: Crocin can be used as an enhancer for neural differentiation of MSCs in vitro in the presence of retinoic acid. The mechanism is proposed through Notch and CREB/BDNF signaling pathways.
■通过分化成骨细胞和脂肪细胞来评估收获的ADSCs的分化能力和干细胞潜能,流式细胞术检测间充质CD标志物的表达。用MTT测定法评估藏红花素的最佳剂量。Crocin,视黄酸,CREB/BDNF,将Notch抑制剂和它们的组合加入培养基中。Jag1,Hes1,Notch,在第7、14和21天通过RT-PCR分析BDNF基因表达,而CREB,DCX,通过免疫荧光分析SOX2和NeuN表达。
■间充质CD标记物的表达以及成脂和成骨分化证实了ADSCs的起源和性质。藏红花素的最佳剂量为1mM。藏红花素显著增加(P<0.05),而抑制剂(DATP和萘酚)显着(P<0.05)降低Jag1,Hes1,Notch,和BDNF表达。免疫荧光评估显示DCX的表达,BDNF,中子,和Sox2蛋白在施用藏红花素后显著升高(P<0.05),在施用抑制剂后显著降低(P<0.05)。
■藏红花素可以用作在视黄酸存在下体外MSCs神经分化的增强剂。该机制是通过Notch和CREB/BDNF信号通路提出的。