脂质体介导NELL1基因对脂肪干细胞影响的体外研究

The effect of NELL1 gene mediated by liposome on the poliferatiaon and differentiation of adipose-derived stem cells in vitro

目的研究Nel样分子I型(nel-like typeⅠmolecular,NELL1)对大鼠脂肪干细胞(adipose-derived stem cells,ADSCs)增殖和分化的影响。方法分离培养大鼠ADSCs并进行传代;在成骨诱导条件下,以脂质体为载体,分别搭载NELL1质粒和增强型绿色荧光蛋白(enhanced green fluorescent protein,EGFP)质粒作为实验组和对照组,转染第三代ADSCs;通过MTT、RT-q PCR、碱性磷酸酶(alkaline phosphatase,ALP)活性以及茜素红染色方法检测细胞增殖和分化能力;采用单因素方差分析进行统计学运算。结果实验组与对照组对比,细胞数量差异无统计学意义(P>0.05),ALP活性及钙结节形成量实验组明显高于对照组。第3d和第7d时,实验组成骨相关基因Runx2、ColⅠ、ALP表达增加(P<0.01),14d时ColⅠm RNA表达增加(P<0.05)。结论 NELL1对ADSCs增殖无明显影响,促进其向成骨细胞分化。

Objective To study the effect of Nel-like type I molecular （NELL1） gene mediated by liposome on the poliferatiaon and osteogenic differentiation of rat adipose-derived stem cells （ADSCs） in vitro. Methods ADSCs were primary cultured and subcuhured in vitro. Liposome was used for gene transfection. The third generation ADSCs were transfected with NELL1 plasmid （experimental group） and EGFP plasmid （control group） respectively, and cultured under osteogenie condition. Mq＇T assay, RT-qPCR, alkaline phosphatase （ALP） activity assay and alizarin red staining were used to test the ability of proliferation and differentiation of the cells. One-way analysis of variance was used to test for statistical significance. Results There was no significant difference in cell proliferation between experimental group and control group （P〉0.05）. Compared with control group, ALP expression and mineralized nodule formation was higher in experiment group （P〈0.01）, and the expression of Runx2, Col I and ALP increased at 3rd and 7th day （P〈0.05）. Conclusion NELL1 may promote the osteoblastic differentiation of ADSCs in cell proliferation in cell proliferation but may be irrelevant to the proliferation.