Jin Jin-lan, Zhuang Han-ping, Wei Jian-rui, Feng Zhi-shun, Deng Zhe-tong, Zhang Min
Central Intensive Care Unit, Guangzhou Red Cross Hospital, Fourth Affiliated Hospital, Jinan University, Guangzhou 510220, Guangdong, China.
Zhongguo Wei Zhong Bing Ji Jiu Yi Xue. 2011 Jun;23(6):355-8.
To research the role of Notch signaling during the differentiation of bone marrow mesenchymal stem cells (MSCs) into endothelial cells and its effect on the functions of the differentiated cells.
Rat bone marrow MSCs were isolated and cultured in vitro, then the cells were treated with vascular endothelial growth factor (VEGF165) and basic fibroblast growth factor (bFGF) for 2 weeks to induce it to differentiate into endothelial cells. The differentiated cells were identified by fluorescence immunoassay. The receptors and ligands of the Notch signaling were detected by reverse transcription-polymerase chain reaction (RT-PCR) before and after the differentiation. γ-secretase inhibitor was used to block Notch pathway. Migration ability of cells were assessed by scarification test. Cells were inoculated on semisolid gel to study their ability of forming the capillary-like structure.
After inducing MSCs to differentiate into endothelial cells by VEGF165 and bFGF, MSCs gained the characteristics of the endothelial cells with expression of CD31 and Flk1. There were Notch1 mRNA and Jagged1 mRNA expressions in rat bone marrow MSCs. The expression changes in the receptor Notch1 were not statistically significant on the differentiated cells (0.59±0.01 vs. 0.59±0.01, P>0.05), but there was a trend towards an increase of Jagged1 mRNA (1.01±0.02 vs. 0.99±0.03, P>0.05). When Notch pathway was blocked, the differentiated cells' migration ability was increased (number of cells on the scratched area: 44.61±4.34 vs. 40.06±2.43, P<0.05), and the ability of forming capillary-like structure was also increased (cells classification: 3.67±0.82 vs. 2.00±0.89, P<0.01).
Notch signaling may have an important role during the differentiation of MSCs into endothelial cells. The function of differentiated cells were strengthened when Notch pathway was blocked.
研究Notch信号通路在骨髓间充质干细胞（MSCs）向内皮细胞分化过程中的作用及其对分化后细胞功能的影响。
分离培养大鼠骨髓间充质干细胞，然后用血管内皮生长因子（VEGF165）和碱性成纤维细胞生长因子（bFGF）处理细胞2周，诱导其分化为内皮细胞。采用荧光免疫分析法鉴定分化后的细胞。在分化前后通过逆转录-聚合酶链反应（RT-PCR）检测Notch信号通路的受体和配体。用γ-分泌酶抑制剂阻断Notch通路。通过划痕试验评估细胞的迁移能力。将细胞接种在半固体凝胶上，研究其形成毛细血管样结构的能力。
经VEGF165和bFGF诱导MSCs分化为内皮细胞后，MSCs获得了内皮细胞的特征，表达CD31和Flk1。大鼠骨髓间充质干细胞中有Notch1 mRNA和Jagged1 mRNA表达。分化后的细胞中受体Notch1的表达变化无统计学意义（0.59±0.01 vs. 0.59±0.01，P>0.05），但Jagged1 mRNA有升高趋势（1.01±0.02 vs. 0.99±0.03，P>0.05）。当Notch通路被阻断时，分化后细胞的迁移能力增强（划痕区域的细胞数：44.61±4.34 vs. 40.06±2.43，P<0.05），形成毛细血管样结构的能力也增强（细胞分级：3.67±0.82 vs. 2.00±0.89，P<0.01）。
Notch信号通路在MSCs向内皮细胞分化过程中可能起重要作用。Notch通路被阻断时，分化后细胞的功能增强。
