Influence of Vascular Endothelial Growth Factor Stimulation and Serum Deprivation on Gene Activation Patterns of Human Adipose Tissue-derived Stromal Cells

Overview

Journal Stem Cell Res Ther

Publisher Biomed Central

Date 2015 Apr 19

PMID 25889587

Citations 18

Authors

Josefine Tratwal

Anders Bruun Mathiasen

Morten Juhl

Sonja Kim Brorsen

Jens Kastrup

Annette Ekblond

Affiliations

Soon will be listed here.

Abstract

Introduction: 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.

Methods: 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.

Results: 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).

Conclusion: 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.

Citing Articles

Apoptosis-induced exosomes from human exfoliated deciduous teeth enhance angiogenesis in human umbilical vein endothelial cells.

Sunartvanichkul T, Chaweewannakorn C, Tabtimmai L, Chiangjong W, Iwasaki K, Pattanapanyasat K Sci Rep. 2024; 14(1):27921.

PMID: 39537956 PMC: 11561270. DOI: 10.1038/s41598-024-79692-6.

Role of Adipose-Derived Mesenchymal Stem Cells in Bone Regeneration.

Lau C, Park S, Ethiraj L, Singh P, Raj G, Quek J Int J Mol Sci. 2024; 25(12).

PMID: 38928517 PMC: 11204188. DOI: 10.3390/ijms25126805.

Advancements in adipose-derived stem cell therapy for skin fibrosis.

Liu Y, Sun J, Ho C, Gao Y, Wen D, Liu Y World J Stem Cells. 2023; 15(5):342-353.

PMID: 37342214 PMC: 10277960. DOI: 10.4252/wjsc.v15.i5.342.

Secretome from human adipose-derived mesenchymal stem cells promotes blood vessel formation and pericyte coverage in experimental skin repair.

Silveira B, Ribeiro T, Freitas R, Carreira A, Goncalves M, Sogayar M PLoS One. 2022; 17(12):e0277863.

PMID: 36534643 PMC: 9762598. DOI: 10.1371/journal.pone.0277863.

Protein biomarkers of cardiac remodeling and inflammation associated with HFpEF and incident events.

Regan J, Truby L, Tahir U, Katz D, Nguyen M, Kwee L Sci Rep. 2022; 12(1):20072.

PMID: 36418363 PMC: 9684116. DOI: 10.1038/s41598-022-24226-1.

References

Tremble P, Lane T, Sage E, Werb Z . SPARC, a secreted protein associated with morphogenesis and tissue remodeling, induces expression of metalloproteinases in fibroblasts through a novel extracellular matrix-dependent pathway. J Cell Biol. 1993; 121(6):1433-44. PMC: 2119706. DOI: 10.1083/jcb.121.6.1433. View

Buerke M, Murohara T, Skurk C, Nuss C, Tomaselli K, Lefer A . Cardioprotective effect of insulin-like growth factor I in myocardial ischemia followed by reperfusion. Proc Natl Acad Sci U S A. 1995; 92(17):8031-5. PMC: 41280. DOI: 10.1073/pnas.92.17.8031. View

Cleutjens J, Kandala J, Guarda E, Guntaka R, Weber K . Regulation of collagen degradation in the rat myocardium after infarction. J Mol Cell Cardiol. 1995; 27(6):1281-92. DOI: 10.1016/s0022-2828(05)82390-9. View

Kuno K, Matsushima K . ADAMTS-1 protein anchors at the extracellular matrix through the thrombospondin type I motifs and its spacing region. J Biol Chem. 1998; 273(22):13912-7. DOI: 10.1074/jbc.273.22.13912. View

Cao Y, Sun Z, Liao L, Meng Y, Han Q, Zhao R . Human adipose tissue-derived stem cells differentiate into endothelial cells in vitro and improve postnatal neovascularization in vivo. Biochem Biophys Res Commun. 2005; 332(2):370-9. DOI: 10.1016/j.bbrc.2005.04.135. View

Zhang B, Kirov S, Snoddy J . WebGestalt: an integrated system for exploring gene sets in various biological contexts. Nucleic Acids Res. 2005; 33(Web Server issue):W741-8. PMC: 1160236. DOI: 10.1093/nar/gki475. View

Cheng L, Guo X, Yang X, Chong M, Cheng J, Li G . Delta-sarcoglycan is necessary for early heart and muscle development in zebrafish. Biochem Biophys Res Commun. 2006; 344(4):1290-9. DOI: 10.1016/j.bbrc.2006.03.234. View

Rocnik E, Liu P, Sato K, Walsh K, Vaziri C . The novel SPARC family member SMOC-2 potentiates angiogenic growth factor activity. J Biol Chem. 2006; 281(32):22855-64. DOI: 10.1074/jbc.M513463200. View

Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D . Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 2006; 8(4):315-7. DOI: 10.1080/14653240600855905. View

10.

van Wijk B, Moorman A, van den Hoff M . Role of bone morphogenetic proteins in cardiac differentiation. Cardiovasc Res. 2006; 74(2):244-55. DOI: 10.1016/j.cardiores.2006.11.022. View

11.

Klapholz-Brown Z, Walmsley G, Nusse Y, Nusse R, Brown P . Transcriptional program induced by Wnt protein in human fibroblasts suggests mechanisms for cell cooperativity in defining tissue microenvironments. PLoS One. 2007; 2(9):e945. PMC: 1976560. DOI: 10.1371/journal.pone.0000945. View

12.

Sadat S, Gehmert S, Song Y, Yen Y, Bai X, Gaiser S . The cardioprotective effect of mesenchymal stem cells is mediated by IGF-I and VEGF. Biochem Biophys Res Commun. 2007; 363(3):674-9. DOI: 10.1016/j.bbrc.2007.09.058. View

13.

Haack-Sorensen M, Friis T, Bindslev L, Mortensen S, Johnsen H, Kastrup J . Comparison of different culture conditions for human mesenchymal stromal cells for clinical stem cell therapy. Scand J Clin Lab Invest. 2007; 68(3):192-203. DOI: 10.1080/00365510701601681. View

14.

Schellings M, Vanhoutte D, Swinnen M, Cleutjens J, Debets J, van Leeuwen R . Absence of SPARC results in increased cardiac rupture and dysfunction after acute myocardial infarction. J Exp Med. 2008; 206(1):113-23. PMC: 2626676. DOI: 10.1084/jem.20081244. View

15.

Lee R, Seo M, Pulin A, Gregory C, Ylostalo J, Prockop D . The CD34-like protein PODXL and alpha6-integrin (CD49f) identify early progenitor MSCs with increased clonogenicity and migration to infarcted heart in mice. Blood. 2008; 113(4):816-26. PMC: 2630267. DOI: 10.1182/blood-2007-12-128702. View

16.

Sanchez C, Oskowitz A, Pochampally R . Epigenetic reprogramming of IGF1 and leptin genes by serum deprivation in multipotential mesenchymal stromal cells. Stem Cells. 2008; 27(2):375-82. PMC: 4943331. DOI: 10.1634/stemcells.2008-0546. View

17.

Dobaczewski M, Gonzalez-Quesada C, Frangogiannis N . The extracellular matrix as a modulator of the inflammatory and reparative response following myocardial infarction. J Mol Cell Cardiol. 2009; 48(3):504-11. PMC: 2824059. DOI: 10.1016/j.yjmcc.2009.07.015. View

18.

Basciano L, Nemos C, Foliguet B, de Isla N, De Carvalho M, Tran N . Long term culture of mesenchymal stem cells in hypoxia promotes a genetic program maintaining their undifferentiated and multipotent status. BMC Cell Biol. 2011; 12:12. PMC: 3073900. DOI: 10.1186/1471-2121-12-12. View

19.

Meng X, Ezzati P, Wilkins J . Requirement of podocalyxin in TGF-beta induced epithelial mesenchymal transition. PLoS One. 2011; 6(4):e18715. PMC: 3075272. DOI: 10.1371/journal.pone.0018715. View

20.

Sassoli C, Pini A, Mazzanti B, Quercioli F, Nistri S, Saccardi R . Mesenchymal stromal cells affect cardiomyocyte growth through juxtacrine Notch-1/Jagged-1 signaling and paracrine mechanisms: clues for cardiac regeneration. J Mol Cell Cardiol. 2011; 51(3):399-408. DOI: 10.1016/j.yjmcc.2011.06.004. View