碱性成纤维细胞生长因子影响脂肪干细胞的血管内皮迁移

Effect of basic fibroblast growth factor on the migration of human adipose-derived stem cells toward vascular endothelium

背景:良好血运机制的建立是工程化组织成功植入的关键。目的:观察外源性碱性成纤维细胞生长因子对植入小鼠皮下的人脂肪来源干细胞-透明质酸钠复合物中人脂肪来源干细胞向血管内皮迁移情况的影响。方法:由吸脂术所得的脂肪组织中分离提取人脂肪来源干细胞进行培养传代,取第3代人脂肪来源干细胞进行cm-dil荧光标记后制成5×109 L-1的细胞悬液,碱性成纤维细胞生长因子配成2 mg/L的工作液。将由0.25 mL透明质酸钠凝胶、0.2 mL细胞悬液和0.05 mL工作液/DMEM混合制成的碱性成纤维细胞生长因子组/对照组移植物分别植入小鼠背部左右两侧皮下作自身对照,6周后取材,行苏木精-伊红染色和免疫荧光标记血管内皮细胞进行观察分析。结果与结论:植入处未出现结节、坏死及液化,取材时无凝胶残留。苏木精-伊红染色见标本性质多为脂肪组织及疏松结缔组织。荧光显微镜下仍可见标记人脂肪来源干细胞的cm-dil荧光,其与标记小鼠血管内皮的FITC荧光重合数碱性成纤维细胞生长因子组多于对照组(P<0.05)。提示碱性成纤维细胞生长因子促进透明质酸钠支架中的人脂肪来源干细胞向血管内皮迁移、分化。

BACKGROUND：The establishment of a good blood supply is a key mechanism for successful implantation of engineered tissues. OBJECTIVE：To observe the effect of basic fibroblast growth factor on the migration of human adipose-derived stem cells via implanting the human adipose-derived stem cells and sodium hyaluronate composite graft at the subcutaneous site of BALB/C mice, in order to explore an optimal scheme for soft tissue reconstruction. METHODS：Human adipose-derived stem cells were isolated from the adipose tissue of healthy cosmetic patients which received liposuction, and the cells were subcultured. Then 5×10^9/L passage 3 cellsuspension labeled by cm-dil was prepared. The working solution containing 2 mg/L basic fibroblast growth factor was prepared. Composite tissue al-lografts which were the mixtures of 0.25 mL sodium hyaluronate, 0.2 mL cellsuspension and 0.05 mL working solution or DMEM were implanted into the subcutaneous site of both sides of the mouse back. Specimens were taken at 6 weeks after operation and were evaluated histological y after hematoxylin-eosin and vascular immunofluorescent staining. RESULTS AND CONCLUSION：No necrosis, liquefaction, nodular tissue or gel remained in operated position. The hematoxylin-eosin staining showed the main components of the specimens were the adipose tissue and the loose connective tissue. The immunofluorescence staining showed the overlaps between the cm-dil fluorescence＆amp;nbsp;from human adipose-derived stem cells and the FITC fluorescence from the vascular endothelium in the experimental group were more than those in the control group （P〈0.05）. Basic fibroblast growth factor promotes the migration and the differentiation of human adipose-derived stem cells in the sodium hyaluronate scaffold into vascular endothelium.