Influence of vascular endothelial growth factor stimulation and serum deprivation on gene activation patterns of human adipose tissue-derived stromal cells.
Influence of vascular endothelial growth factor stimulation and serum deprivation on gene activation patterns of human adipose tissue-derived stromal cells.
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血管内皮生长因子刺激和血清缺乏对人脂肪组织衍生基质细胞基因激活模式的影响。
DOI：
10.1186/s13287-015-0062-9
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发表时间：
2015-04-13
影响因子：
7.5
通讯作者：
Ekblond A
中科院分区：
文献类型：
--
作者：
Tratwal J;Mathiasen AB;Juhl M;Brorsen SK;Kastrup J;Ekblond A
血管内皮生长因子（VEGF）刺激间充质基质细胞和脂肪组织源性基质细胞（ASCs）已在多个动物研究和临床试验中用于再生目的。VEGF刺激被认为可以促进血管生成，而VEGF刺激通常在血清剥夺的情况下进行。潜在的再生分子机制有很多，影响因素的作用还不确定。本研究旨在探讨体外血清剥夺和VEGF刺激对ASCs基因表达模式的影响。比较完全培养基培养的ASCs、无血清培养基培养的ASCs和无血清培养基中VEGF刺激的ASCs的基因表达情况。根据国际细胞治疗学会制定的标准，流式细胞术证实ASC特征。使用Affymetrix HT HG-U133+ GeneChip®获得微阵列基因表达。使用京都基因与基因组百科全书和基因本体术语进行基因集富集分析。选择感兴趣的基因转录通过定量PCR证实。与完全培养基中的ASCs相比，血清剥夺和血清剥夺联合VEGF分别使190个和108个基因发生显著改变。血清剥夺ASCs与血清剥夺ASCs联合VEGF刺激的基因表达模式无显著差异。在这两种情况下，最显著上调的基因是生长因子（IGF1、BMP6、PDGFD、FGF9）、粘附分子CLSTN2、细胞外基质相关蛋白（如基质蛋白SMOC2、SPON1和ADAMTS12）和增殖抑制剂（JAG1）。最显著下调的基因包括基质金属蛋白酶（MMP3、MMP1）、增殖标志物（CDKN3）和GREM2 （BMP6拮抗剂）。ASC基因表达变化的决定性因素是血清饥饿而非VEGF刺激。作为使用方案的主要结果，生长因子、基质细胞蛋白和基质金属蛋白酶协同表达的变化与直接的促血管生成旁分泌机制不同。体外血清饥饿（无论是否存在VEGF）似乎有利于心肌保护、细胞外基质重塑和血管成熟，这与梗死愈合的晚期成熟阶段有关。
Stimulation of mesenchymal stromal cells and adipose tissue-derived stromal cells (ASCs) with vascular endothelial growth factor (VEGF) has been used in multiple animal studies and clinical trials for regenerative purposes. VEGF stimulation is believed to promote angiogenesis and VEGF stimulation is usually performed under serum deprivation. Potential regenerative molecular mechanisms are numerous and the role of contributing factors is uncertain. The aim of the current study was to investigate the effect of in vitro serum deprivation and VEGF stimulation on gene expression patterns of ASCs. Gene expressions of ASCs cultured in complete medium, ASCs cultured in serum-deprived medium and ASCs stimulated with VEGF in serum-deprived medium were compared. ASC characteristics according to criteria set by the International Society of Cellular Therapy were confirmed by flow cytometry. Microarray gene expressions were obtained using the Affymetrix HT HG-U133+ GeneChip®. Gene set enrichment analysis was performed using the Kyoto Encyclopedia of Genes and Genomes and gene ontology terms. Transcription of selected genes of interest was confirmed by quantitative PCR. Compared to ASCs in complete medium, 190 and 108 genes were significantly altered by serum deprivation and serum deprivation combined with VEGF, respectively. No significant differences in gene expression patterns between serum-deprived ASCs and serum-deprived ASCs combined with VEGF stimulation were found. Genes most prominently and significantly upregulated by both conditions were growth factors (IGF1, BMP6, PDGFD, FGF9), adhesion molecule CLSTN2, extracellular matrix-related proteins such as matricellular proteins SMOC2, SPON1 and ADAMTS12, and inhibitors of proliferation (JAG1). The most significantly downregulated genes included matrix metalloproteinases (MMP3, MMP1), and proliferation markers (CDKN3) and GREM2 (a BMP6 antagonist). The decisive factor for the observed change in ASC gene expression proves to be serum starvation rather than VEGF stimulation. Changes in expression of growth factors, matricellular proteins and matrix metalloproteinases in concert, diverge from direct pro-angiogenic paracrine mechanisms as a primary consequence of the used protocol. In vitro serum starvation (with or without VEGF present) appears to favour cardioprotection, extracellular matrix remodelling and blood vessel maturation relevant for the late maturation phase in infarct healing.
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影响因子：
14.9
作者：
Kolesnikov N;Hastings E;Keays M;Melnichuk O;Tang YA;Williams E;Dylag M;Kurbatova N;Brandizi M;Burdett T;Megy K;Pilicheva E;Rustici G;Tikhonov A;Parkinson H;Petryszak R;Sarkans U;Brazma A
通讯作者：
Brazma A
影响因子：
12.3
作者：
Gentleman RC;Carey VJ;Bates DM;Bolstad B;Dettling M;Dudoit S;Ellis B;Gautier L;Ge Y;Gentry J;Hornik K;Hothorn T;Huber W;Iacus S;Irizarry R;Leisch F;Li C;Maechler M;Rossini AJ;Sawitzki G;Smith C;Smyth G;Tierney L;Yang JY;Zhang J
通讯作者：
Zhang J
影响因子：
8.5
作者：
Chua, K. H.;Raduan, F.;Sathapan, S.
通讯作者：
Sathapan, S.
影响因子：
6.7
作者：
Cabrita, MA;Christofori, G
通讯作者：
Christofori, G
DOI：
10.1080/00365510701601681
发表时间：
2008-01-01
影响因子：
2.1
作者：
Haack-Sorensen, M.;Friis, T.;Kastrup, J.
通讯作者：
Kastrup, J.