It has been demonstrated that human adipose tissue-derived mesenchymal stem cells (hASCs) enhance vascular density in ischemic tissues, suggesting that they can differentiate into vascular cells or release angiogenic factors that may stimulate neoangiogenesis. Moreover, there is evidence that shear stress (SS) may activate proliferation and differentiation of embryonic and endothelial precursor stem cells into endothelial cells (ECs). In this work, we investigated the effect of laminar SS in promoting differentiation of hASCs into ECs. SS (10 dyn/cm2 up to 96 h), produced by a cone plate system, failed to induce EC markers (CD31, vWF, Flk-1) on hASC assayed by RT-PCR and flow cytometry. In contrast, there was a cumulative production of nitric oxide (determined by Griess Reaction) and vascular endothelial growth factor (VEGF; by ELISA) up to 96 h of SS stimulation ( in nmol/104 cells: static: 0.20 ± 0.03; SS: 1.78 ± 0.38, n = 6; VEGF in pg/104 cells: static: 191.31 ± v35.29; SS: 372.80 ± 46.74, n = 6, P < 0.05). Interestingly, the VEGF production was abrogated by 5 mM N(G)-l-nitro-arginine methyl ester (l-NAME) treatment (VEGF in pg/104 cells: SS: 378.80 ± 46.74, n = 6; SS + l-NAME: 205.84 ± 91.66, n = 4, P < 0.05). The results indicate that even though SS failed to induce EC surface markers in hASC under the tested conditions, it stimulated NO-dependent VEGF production.
Get full access to this article
View all access options for this article.
References
1.
RihaGM, LinPH, LumsdenAB, YaoQ and ChenC. (2005). Roles of hemodynamic forces in vascular cell differentiation. Ann Biomed Eng, 33:772–779.
2.
RubanyiGM, FreayAD, KauserK, JohnsA and HarderDR. (1990). Mechanoreception by the endothelium: mediators and mechanisms of pressure- and flow-induced vascular responses. Blood Vessels, 27:246–257.
3.
WangH, RihaGM, YanS, LiM, ChaiH, YangH, YaoQ and ChenC. (2005). Shear stress induces endothelial differentiation from a murine embryonic mesenchymal progenitor cell line. Arterioscler Thromb Vasc Biol, 25:1817–1823.
4.
YamamotoK, SokabeT, WatabeT, MiyazonoK, YamashitaJK, ObiS, OhuraN, MatsushitaA, KamiyaA and AndoJ. (2005). Fluid shear stress induces differentiation of Flk-1-positive embryonic stem cells into vascular endothelial cells in vitro. Am J Physiol Heart Circ Physiol, 288:H1915–H1924.
5.
YamamotoK, TakahashiT, AsaharaT, OhuraN, SokabeT, KamiyaA and AndoJ. (2003). Proliferation, differentiation, and tube formation by endothelial progenitor cells in response to shear stress. J Appl Physiol, 95:2081–2088.
6.
WuCC, ChaoYC, ChenCN, ChienS, ChenYC, ChienCC, ChiuJJ and Linju YenB. (2008). Synergism of biochemical and mechanical stimuli in the differentiation of human placenta-derived multipotent cells into endothelial cells. J Biomech, 41:813–821.
7.
WangH, YanS, ChaiH, RihaGM, LiM, YaoQ and ChenC. (2006). Shear stress induces endothelial transdifferentiation from mouse smooth muscle cells. Biochem Biophys Res Commun, 346:860–865.
8.
Planat-BenardV, SilvestreJS, CousinB, AndreM, NibbelinkM, TamaratR, ClergueM, MannevilleC, Saillan-BarreauC, DuriezM, TedguiA, LevyB, PenicaudL and CasteillaL. (2004). Plasticity of human adipose lineage cells toward endothelial cells: physiological and therapeutic perspectives. Circulation, 109:656–663.
9.
MiranvilleA, HeeschenC, SengenesC, CuratCA, BusseR and BouloumieA. (2004). Improvement of postnatal neovascularization by human adipose tissue-derived stem cells. Circulation, 110:349–355.
10.
MoonMH, KimSY, KimYJ, KimSJ, LeeJB, BaeYC, SungSM and JungJS. (2006). Human adipose tissue-derived mesenchymal stem cells improve postnatal neovascularization in a mouse model of hindlimb ischemia. Cell Physiol Biochem, 17:279–290.
11.
CaoY, SunZ, LiaoL, MengY, HanQ and ZhaoRC. (2005). Human adipose tissue-derived stem cells differentiate into endothelial cells in vitro and improve postnatal neovascularization in vivo. Biochem Biophys Res Commun, 332:370–379.
12.
CaoY, MengY, SunZ, LiaoLM, HanQ, LiJ, LiuYN and ZhaoCH. (2005). [Potential of human adipose tissue derived adult stem cells differentiate into endothelial cells]. Zhongguo Yi Xue Ke Xue Yuan Xue Bao, 27:678–682.
13.
LozitoTP, KuoCK, TaboasJM and TuanRS. (2009). Human mesenchymal stem cells express vascular cell phenotypes upon interaction with endothelial cell matrix. J Cell Biochem, 107:714–722.
14.
ZukPA, ZhuM, AshjianP, De UgarteDA, HuangJI, MizunoH, AlfonsoZC, FraserJK, BenhaimP and HedrickMH. (2002). Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell, 13:4279–4295.
15.
BlandeIS, BassanezeV, Lavini-RamosC, FaeKC, KalilJ, MiyakawaAA, SchettertIT and KriegerJE. (2009). Adipose tissue mesenchymal stem cell expansion in animal serum-free medium supplemented with autologous human platelet lysate. Transfusion. [Epub ahead of print]
16.
MalekA and IzumoS. (1992). Physiological fluid shear stress causes downregulation of endothelin-1 mRNA in bovine aortic endothelium. Am J Physiol, 263:C389–C396.
17.
MiyakawaAA, de Lourdes JunqueiraM and KriegerJE. (2004). Identification of two novel shear stress responsive elements in rat angiotensin I converting enzyme promoter. Physiol Genomics, 17:107–113.
18.
DingAH, NathanCF and StuehrDJ. (1988). Release of reactive nitrogen intermediates and reactive oxygen intermediates from mouse peritoneal macrophages. Comparison of activating cytokines and evidence for independent production. J Immunol, 141:2407–2412.
19.
AbramoffMD, MagelhaesPJ and RamSJ. (2004). Image Processing with ImageJ. Biophotonics Int, 11:36–42.
20.
LeeRH, KimB, ChoiI, KimH, ChoiHS, SuhK, BaeYC and JungJS. (2004). Characterization and expression analysis of mesenchymal stem cells from human bone marrow and adipose tissue. Cell Physiol Biochem, 14:311–324.
21.
BerkBC, CorsonMA, PetersonTE and TsengH. (1995). Protein kinases as mediators of fluid shear stress stimulated signal transduction in endothelial cells: a hypothesis for calcium-dependent and calcium-independent events activated by flow. J Biomech, 28:1439–1450.
22.
UematsuM, OharaY, NavasJP, NishidaK, MurphyTJ, AlexanderRW, NeremRM and HarrisonDG. (1995). Regulation of endothelial cell nitric oxide synthase mRNA expression by shear stress. Am J Physiol, 269:C1371–C1378.
23.
KimuraH, OguraT, KurashimaY, WeiszA and EsumiH. (2002). Effects of nitric oxide donors on vascular endothelial growth factor gene induction. Biochem Biophys Res Commun, 296:976–982.
24.
SasakiT, YoshidaK, KondoH, OhmoriH and KuniyasuH. (2005). Heme oxygenase-1 accelerates protumoral effects of nitric oxide in cancer cells. Virchows Arch, 446:525–531.
25.
AzumaN, AkasakaN, KitoH, IkedaM, GahtanV, SasajimaT and SumpioBE. (2001). Role of p38 MAP kinase in endothelial cell alignment induced by fluid shear stress. Am J Physiol Heart Circ Physiol, 280:H189–H197.
26.
ZengL, XiaoQ, MargaritiA, ZhangZ, ZampetakiA, PatelS, CapogrossiMC, HuY and XuQ. (2006). HDAC3 is crucial in shear- and VEGF-induced stem cell differentiation toward endothelial cells. J Cell Biol, 174:1059–1069.
JonesEA, le NobleF and EichmannA. (2006). What determines blood vessel structure? Genetic prespecification vs. hemodynamics. Physiology (Bethesda), 21:388–395.
29.
StolbergS and McCloskeyKE. (2009). Can shear stress direct stem cell fate?. Biotechnol Prog, 25:10–19.
30.
ZhangP, BaxterJ, VinodK, TulenkoTN and DimuzioP. (2009). Endothelial Differentiation of amniotic fluid-derived stem cells: synergism of biochemical and shear force stimuli. Stem Cells Dev. [Epub ahead of print]
31.
MooreMA. (2002). Putting the neo into neoangiogenesis. J Clin Invest, 109:313–315.
32.
CleaverO and KriegPA. (1998). VEGF mediates angioblast migration during development of the dorsal aorta in Xenopus. Development, 125:3905–3914.
33.
ChoiK, KennedyM, KazarovA, PapadimitriouJC and KellerG. (1998). A common precursor for hematopoietic and endothelial cells. Development, 125:725–732.
34.
Martinez-EstradaOM, Munoz-SantosY, JulveJ, ReinaM and VilaroS. (2005). Human adipose tissue as a source of Flk-1+ cells: new method of differentiation and expansion. Cardiovasc Res, 65:328–333.
35.
MorozLL, NorbySW, CruzL, SweedlerJV, GilletteR and ClarksonRB. (1998). Non-enzymatic production of nitric oxide (NO) from NO synthase inhibitors. Biochem Biophys Res Commun, 253:571–576.
36.
MaisA, KleinT, UllrichV, SchudtC and LauerG. (2006). Prostanoid pattern and iNOS expression during chondrogenic differentiation of human mesenchymal stem cells. J Cell Biochem, 98:798–809.
37.
SatoK, OzakiK, OhI, MeguroA, HatanakaK, NagaiT, MuroiK and OzawaK. (2007). Nitric oxide plays a critical role in suppression of T-cell proliferation by mesenchymal stem cells. Blood, 109:228–234.
38.
KnippenbergM, HelderMN, DoulabiBZ, SemeinsCM, WuismanPI and Klein-NulendJ. (2005). Adipose tissue-derived mesenchymal stem cells acquire bone cell-like responsiveness to fluid shear stress on osteogenic stimulation. Tissue Eng, 11:1780–1788.
39.
TjabringaGS, VezeridisPS, Zandieh-DoulabiB, HelderMN, WuismanPI and Klein-NulendJ. (2006). Polyamines modulate nitric oxide production and COX-2 gene expression in response to mechanical loading in human adipose tissue-derived mesenchymal stem cells. Stem Cells, 24:2262–2269.
40.
DimmelerS, FlemingI, FisslthalerB, HermannC, BusseR and ZeiherAM. (1999). Activation of nitric oxide synthase in endothelial cells by Akt-dependent phosphorylation. Nature, 399:601–605.
41.
EmingSA and KriegT. (2006). Molecular mechanisms of VEGF-A action during tissue repair. J Investig Dermatol Symp Proc, 11:79–86.
42.
KimuraH and EsumiH. (2003). Reciprocal regulation between nitric oxide and vascular endothelial growth factor in angiogenesis. Acta Biochim Pol, 50:49–59.
43.
WangY, CrisostomoPR, WangM, WeilB, AbarbanellA, PoynterJ, ManukyanMC and MeldrumDR. (2008). Nitric oxide suppresses the secretion of vascular endothelial growth factor and hepatocyte growth factor from human mesenchymal stem cells. Shock, 30:527–531.
44.
RehmanJ, TraktuevD, LiJ, Merfeld-ClaussS, Temm-GroveCJ, BovenkerkJE, PellCL, JohnstoneBH, ConsidineRV and MarchKL. (2004). Secretion of angiogenic and antiapoptotic factors by human adipose stromal cells. Circulation, 109:1292–1298.
45.
KimuraH, WeiszA, KurashimaY, HashimotoK, OguraT, D’AcquistoF, AddeoR, MakuuchiM and EsumiH. (2000). Hypoxia response element of the human vascular endothelial growth factor gene mediates transcriptional regulation by nitric oxide: control of hypoxia-inducible factor-1 activity by nitric oxide. Blood, 95:189–197.
46.
KomoriK, TsujimuraA, TakaoT, MatsuokaY, MiyagawaY, TakadaS, NonomuraN and OkuyamaA. (2008). Nitric oxide synthesis leads to vascular endothelial growth factor synthesis via the NO/cyclic guanosine 3′,5′-monophosphate (cGMP) pathway in human corpus cavernosal smooth muscle cells. J Sex Med, 5:1623–1635.
47.
TsurumiY, MuroharaT, KrasinskiK, ChenD, WitzenbichlerB, KearneyM, CouffinhalT and IsnerJM. (1997). Reciprocal relation between VEGF and NO in the regulation of endothelial integrity. Nat Med, 3:879–886.
48.
DulakJ, JozkowiczA, Dembinska-KiecA, GuevaraI, ZdzienickaA, Zmudzinska-GrochotD, FlorekI, WojtowiczA, SzubaA and CookeJP. (2000). Nitric oxide induces the synthesis of vascular endothelial growth factor by rat vascular smooth muscle cells. Arterioscler Thromb Vasc Biol, 20:659–666.
49.
JozkowiczA, CookeJP, GuevaraI, HukI, FunovicsP, PachingerO, WeidingerF and DulakJ. (2001). Genetic augmentation of nitric oxide synthase increases the vascular generation of VEGF. Cardiovasc Res, 51:773–783.
50.
DulakJ and JozkowiczA. (2003). Regulation of vascular endothelial growth factor synthesis by nitric oxide: facts and controversies. Antioxid Redox Signal, 5:123–132.
51.
GonçalvesGA, VassalloPF, dos SantosL, SchettertIT, NakamutaJS, BeckerC, TucciPJF and KriegerJK. Intramyocardial transplantation of fibroblasts expressing vascular endothelial growth factor attenuates cardiac dysfunction. Gene Therapy, (in press)
52.
BeckerC, LacchiniS, MuotriAR, da SilvaGJ, CastelliJB, VassalloPF, MenckCF and KriegerJE. (2006). Skeletal muscle cells expressing VEGF induce capillary formation and reduce cardiac injury in rats. Int J Cardiol, 113:348–354.
53.
TogelF, HuZ, WeissK, IsaacJ, LangeC and WestenfelderC. (2005). Administered mesenchymal stem cells protect against ischemic acute renal failure through differentiation-independent mechanisms. Am J Physiol Renal Physiol, 289:F31–F42.
54.
GuoJ, LinGS, BaoCY, HuZM and HuMY. (2007). Anti-inflammation role for mesenchymal stem cells transplantation in myocardial infarction. Inflammation, 30:97–104.
55.
UccelliA, PistoiaV and MorettaL. (2007). Mesenchymal stem cells: a new strategy for immunosuppression?. Trends Immunol, 28:219–226.
56.
GnecchiM, HeH, NoiseuxN, LiangOD, ZhangL, MorelloF, MuH, MeloLG, PrattRE, IngwallJS and DzauVJ. (2006). Evidence supporting paracrine hypothesis for Akt-modified mesenchymal stem cell-mediated cardiac protection and functional improvement. FASEB J, 20:661–669.
