Effects of serum reduction and VEGF supplementation on angiogenic potential of human adipose stromal cells in vitro
Effects of serum reduction and VEGF supplementation on angiogenic potential of human adipose stromal cells in vitro
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DOI：
10.1111/cpr.12029
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发表时间：
2013-06-01
影响因子：
8.5
通讯作者：
Sathapan, S.
中科院分区：
文献类型：
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作者：
Chua, K. H.;Raduan, F.;Sathapan, S.
目的探讨降低血清浓度和血管内皮生长因子(VEGF)对脂肪基质细胞(ASCs)血管生成潜能的影响。材料与方法脂肪基质细胞在三种不同的培养液中培养：(1)F12/DMEM(FD)，从第0代(P0)到P6，添加10%FBS；(2)在P6，FD+2%FBS；(3)在P6，FD+2%FBS+50 ng/mlVEGF。记录ASCs的形态变化和生长速度。用实时定量聚合酶链式反应分析茎、血管生成和内源性基因表达的变化。结果脂肪基质细胞在10%胎牛血清中培养时由成纤维细胞样细胞形态转变为多角形，在2%胎牛血清+血管内皮细胞生长因子培养液中培养。在2%的FBS培养液中，细胞生长速度较慢，但加入血管内皮细胞生长因子后，细胞生长速度加快。实时荧光定量聚合酶链式反应显示ASCs在P1、P5和P6的10%FBS中保持了大部分的干性和血管生成基因的表达，而在P6时在2%的FBS中ASCs的表达显著增加。PECAM-1、VE chaderin、VEGFR-2等内源性基因在10%胎牛血清中连续传代后表达降低，而在2%胎牛血清中传代后表达显著增加。加入血管内皮细胞生长因子后，基因表达水平无明显变化。结论脂肪基质细胞在低血清培养条件下具有较大的血管生成潜能。在添加2%胎牛血清的培养液中，血管内皮生长因子不能增强其血管生成能力。
Objectives This study investigated effects of reduced serum condition and vascular endothelial growth factor (VEGF) on angiogenic potential of adipose stromal cells (ASCs) in vitro. Materials and methods Adipose stromal cells were cultured in three different types of medium: (i) F12/DMEM (FD) supplemented with 10% FBS from passage 0 (P0) to P6; (ii) FD supplemented with 2% FBS at P6; and (iii) FD supplemented with 2% FBS plus 50ng/ml of VEGF at P6. Morphological changes and growth rate of ASCs were recorded. Changes in stemness, angiogenic and endogenic genes' expressions were analysed using Real-Time PCR. Results Adipose stromal cells changed from fibroblast-like shape when cultured in 10% FBS medium to polygonal when cultured in 2% FBS plus VEGF-supplemented medium. Their growth rate was lower in 2% FBS medium, but increased with addition of VEGF. Real-Time PCR showed that ASCs maintained most of their stemness and angiogenic genes' expression in 10% FBS at P1, P5 and P6, but this increased significantly in 2% FBS at P6. Endogenic genes expression such as PECAM-1, VE chaderin and VEGFR-2 decreased after serial passage in 10% FBS, but increased significantly at P6 in 2% FBS. Addition of VEGF did not cause any significant change in gene expression level. Conclusion Adipose stromal cells had greater angiogenic potential when cultured in reduced serum conditions. VEGF did not enhance their angiogenic potential in 2% FBS-supplemented medium.