Mesenchymal stem cells (MSCs) show great promise in blood vessel restoration and vascularization enhancement in many therapeutic situations. Typically, the co-implantation of MSCs with vascular endothelial cells (ECs) is effective for the induction of functional vascularization in vivo, indicating its potential applications in regenerative medicine. The effects of MSCs-ECs-induced vascularization can be modeled in vitro, providing simplified models for understanding their underlying communication. In this article, a contact coculture model in vitro and an RNA-seq approach were employed to reveal the active crosstalk between MSCs and ECs within a short time period at both morphological and transcriptional levels. The RNA-seq results suggested that angiogenic genes were significantly induced upon coculture, and this prevascularization commitment might require the NF-κB signaling. NF-κB blocking and interleukin (IL) neutralization experiments demonstrated that MSCs potentially secreted IL factors including IL1β and IL6 to modulate NF-κB signaling and downstream chemokines during coculture. Conversely, RNA-seq results indicated that the MSCs were regulated by the coculture environment to a smooth muscle commitment within this short period, which largely induced myocardin, the myogenic co-transcriptional factor. These findings demonstrate the mutual molecular mechanism of MSCs-ECs-induced prevascularization commitment in a quick response.
Get full access to this article
View all access options for this article.
References
1.
AbedinM, TintutY and DemerLL. (2004). Mesenchymal stem cells and the artery wall. Circ Res, 95:671–676.
2.
WilliamsAR and HareJM. (2011). Mesenchymal stem cells: biology, pathophysiology, translational findings, and therapeutic implications for cardiac disease. Circ Res, 109:923–940.
3.
PontikoglouC, DeschaseauxF, SensebeL and PapadakiHA. (2011). Bone marrow mesenchymal stem cells: biological properties and their role in hematopoiesis and hematopoietic stem cell transplantation. Stem Cell Rev, 7:569–589.
4.
PatiS, KhakooAY, ZhaoJ, JimenezF, GerberMH, HartingM, RedellJB, GrillR, MatsuoY, et al. (2011). Human mesenchymal stem cells inhibit vascular permeability by modulating vascular endothelial cadherin/beta-catenin signaling. Stem Cells Dev, 20:89–101.
5.
DuffyGP, AhsanT, O'BrienT, BarryF and NeremRM. (2009). Bone marrow-derived mesenchymal stem cells promote angiogenic processes in a time- and dose-dependent manner in vitro. Tissue Eng Part A, 15:2459–2470.
6.
AugerFA, GibotL and LacroixD. (2013). The pivotal role of vascularization in tissue engineering. Annu Rev Biomed Eng, 15:177–200.
7.
LovettM, LeeK, EdwardsA and KaplanDL. (2009). Vascularization strategies for tissue engineering. Tissue Eng Part B Rev, 15:353–370.
8.
KoikeN, FukumuraD, GrallaO, AuP, SchechnerJS and JainRK. (2004). Tissue engineering: creation of long-lasting blood vessels. Nature, 428:138–139.
9.
WangZZ, AuP, ChenT, ShaoY, DaheronLM, BaiH, ArzigianM, FukumuraD, JainRK and ScaddenDT. (2007). Endothelial cells derived from human embryonic stem cells form durable blood vessels in vivo. Nat Biotechnol, 25:317–318.
10.
SamuelR, DaheronL, LiaoS, VardamT, KamounWS, BatistaA, BueckerC, SchaferR, HanX, et al. (2013). Generation of functionally competent and durable engineered blood vessels from human induced pluripotent stem cells. Proc Natl Acad Sci U S A, 110:12774–12779.
11.
SalehFA, WhyteM, AshtonP and GeneverPG. (2011). Regulation of mesenchymal stem cell activity by endothelial cells. Stem Cells Dev, 20:391–403.
12.
KaiglerD, KrebsbachPH, WestER, HorgerK, HuangYC and MooneyDJ. (2005). Endothelial cell modulation of bone marrow stromal cell osteogenic potential. FASEB J, 19:665–667.
13.
BidarraSJ, BarriasCC, BarbosaMA, SoaresR, AmedeeJ and GranjaPL. (2011). Phenotypic and proliferative modulation of human mesenchymal stem cells via crosstalk with endothelial cells. Stem Cell Res, 7:186–197.
14.
BallSG, ShuttleworthAC and KieltyCM. (2004). Direct cell contact influences bone marrow mesenchymal stem cell fate. Int J Biochem Cell Biol, 36:714–727.
15.
TomaC, PittengerMF, CahillKS, ByrneBJ and KesslerPD. (2002). Human mesenchymal stem cells differentiate to a cardiomyocyte phenotype in the adult murine heart. Circulation, 105:93–98.
16.
TsujiH, MiyoshiS, IkegamiY, HidaN, AsadaH, TogashiI, SuzukiJ, SatakeM, NakamizoH, et al. (2010). Xenografted human amniotic membrane-derived mesenchymal stem cells are immunologically tolerated and transdifferentiated into cardiomyocytes. Circ Res, 106:1613–1623.
17.
AuP, TamJ, FukumuraD and JainRK. (2008). Bone marrow-derived mesenchymal stem cells facilitate engineering of long-lasting functional vasculature. Blood, 111:4551–4558.
18.
GrellierM, BordenaveL and AmedeeJ. (2009). Cell-to-cell communication between osteogenic and endothelial lineages: implications for tissue engineering. Trends Biotechnol, 27:562–571.
19.
ChenDY, WeiHJ, LinKJ, HuangCC, WangCC, WuCT, ChaoKT, ChenKJ, ChangY and SungHW. (2013). Three-dimensional cell aggregates composed of HUVECs and cbMSCs for therapeutic neovascularization in a mouse model of hindlimb ischemia. Biomaterials, 34:1995–2004.
20.
LeeWY, TsaiHW, ChiangJH, HwangSM, ChenDY, HsuLW, HungYW, ChangY and SungHW. (2011). Core-shell cell bodies composed of human cbMSCs and HUVECs for functional vasculogenesis. Biomaterials, 32:8446–8455.
21.
LiH, DaculsiR, GrellierM, BareilleR, BourgetC, RemyM and AmedeeJ. (2011). The role of vascular actors in two dimensional dialogue of human bone marrow stromal cell and endothelial cell for inducing self-assembled network. PLoS One, 6:e16767
22.
GrellierM, Ferreira-TojaisN, BourgetC, BareilleR, GuillemotF and AmedeeJ. (2009). Role of vascular endothelial growth factor in the communication between human osteoprogenitors and endothelial cells. J Cell Biochem, 106:390–398.
23.
CarrionB, KongYP, KaiglerD and PutnamAJ. (2013). Bone marrow-derived mesenchymal stem cells enhance angiogenesis via their alpha6beta1 integrin receptor. Exp Cell Res, 319:2964–2976.
24.
Melero-MartinJM, De ObaldiaME, KangSY, KhanZA, YuanL, OettgenP and BischoffJ. (2008). Engineering robust and functional vascular networks in vivo with human adult and cord blood-derived progenitor cells. Circ Res, 103:194–202.
25.
Kunz-SchughartLA, SchroederJA, WondrakM, van ReyF, LehleK, HofstaedterF and WheatleyDN. (2006). Potential of fibroblasts to regulate the formation of three-dimensional vessel-like structures from endothelial cells in vitro. Am J Physiol Cell Physiol, 290:C1385–C1398.
26.
LiH, XueK, KongN, LiuK and ChangJ. (2014). Silicate bioceramics enhanced vascularization and osteogenesis through stimulating interactions between endothelia cells and bone marrow stromal cells. Biomaterials, 35:3803–3818.
27.
LuuNT, McGettrickHM, BuckleyCD, NewsomeP, EdRG, FramptonJ and NashGB. (2013). Crosstalk between mesenchymal stem cells and endothelial cells leads to down-regulation of cytokine-induced leukocyte recruitment. Stem Cells, 31:2690–2702.
28.
MatsushitaT, KibayashiT, KatayamaT, YamashitaY, SuzukiS, KawamataJ, HonmouO, MinamiM and ShimohamaS. (2011). Mesenchymal stem cells transmigrate across brain microvascular endothelial cell monolayers through transiently formed inter-endothelial gaps. Neurosci Lett, 502:41–45.
29.
TeoGS, AnkrumJA, MartinelliR, BoettoSE, SimmsK, SciutoTE, DvorakAM, KarpJM and CarmanCV. (2012). Mesenchymal stem cells transmigrate between and directly through tumor necrosis factor-alpha-activated endothelial cells via both leukocyte-like and novel mechanisms. Stem Cells, 30:2472–2486.
30.
TrapnellC, RobertsA, GoffL, PerteaG, KimD, KelleyDR, PimentelH, SalzbergSL, RinnJL and PachterL. (2012). Differential gene and transcript expression analysis of RNA-seq experiments with TopHat and Cufflinks. Nat Protoc, 7:562–578.
31.
HuangDW, ShermanBT and LempickiRA. (2009). Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc, 4:44–57.
32.
AguirreA, PlanellJA and EngelE. (2010). Dynamics of bone marrow-derived endothelial progenitor cell/mesenchymal stem cell interaction in co-culture and its implications in angiogenesis. Biochem Biophys Res Commun, 400:284–291.
33.
TsudaS, OhtsuruA, YamashitaS, KanetakeH and KandaS. (2002). Role of c-Fyn in FGF-2-mediated tube-like structure formation by murine brain capillary endothelial cells. Biochem Biophys Res Commun, 290:1354–1360.
34.
MochizukiY, NakamuraT, KanetakeH and KandaS. (2002). Angiopoietin 2 stimulates migration and tube-like structure formation of murine brain capillary endothelial cells through c-Fes and c-Fyn. J Cell Sci, 115:175–183.
35.
ZacharekA, ChenJ, CuiX, LiA, LiY, RobertsC, FengY, GaoQ and ChoppM. (2007). Angiopoietin1/Tie2 and VEGF/Flk1 induced by MSC treatment amplifies angiogenesis and vascular stabilization after stroke. J Cereb Blood Flow Metab, 27:1684–1691.
36.
OguraH, MurakamiM, OkuyamaY, TsuruokaM, KitabayashiC, KanamotoM, NishiharaM, IwakuraY and HiranoT. (2008). Interleukin-17 promotes autoimmunity by triggering a positive-feedback loop via interleukin-6 induction. Immunity, 29:628–636.
37.
PugazhenthiS, ZhangY, BouchardR and MahaffeyG. (2013). Induction of an inflammatory loop by interleukin-1beta and tumor necrosis factor-alpha involves NF-kB and STAT-1 in differentiated human neuroprogenitor cells. PLoS One, 8:e69585
38.
PahlHL. (1999). Activators and target genes of Rel/NF-kappa B transcription factors. Oncogene, 18:6853–6866.
39.
DimbergA. (2010). Chemokines in angiogenesis. In: The Chemokine System in Experimental and Clinical Hematology. Springer Berlin Heidelberg, pp 59–80.
WeiL, ZhouW, CroissantJD, JohansenFE, PrywesR, BalasubramanyamA and SchwartzRJ. (1998). RhoA signaling via serum response factor plays an obligatory role in myogenic differentiation. J Biol Chem, 273:30287–30294.
42.
JeonES, ParkWS, LeeMJ, KimYM, HanJ and KimJH. (2008). A Rho kinase/myocardin-related transcription factor-A-dependent mechanism underlies the sphingosylphosphorylcholine-induced differentiation of mesenchymal stem cells into contractile smooth muscle cells. Circ Res, 103:635–642.
43.
ChenS and LechleiderRJ. (2004). Transforming growth factor-beta-induced differentiation of smooth muscle from a neural crest stem cell line. Circ Res, 94:1195–1202.
FuchsS, HofmannA and KirkpatrickC. (2007). Microvessel-like structures from outgrowth endothelial cells from human peripheral blood in 2-dimensional and 3-dimensional co-cultures with osteoblastic lineage cells. Tissue Eng, 13:2577–2588.
46.
BaigueraS and RibattiD. (2013). Endothelialization approaches for viable engineered tissues. Angiogenesis, 16:1–14.
47.
HofmannA, RitzU, VerrierS, EglinD, AliniM, FuchsS, KirkpatrickCJ and RommensPM. (2008). The effect of human osteoblasts on proliferation and neo-vessel formation of human umbilical vein endothelial cells in a long-term 3D co-culture on polyurethane scaffolds. Biomaterials, 29:4217–4226.
48.
EgorovaAD, DeRuiterMC, de BoerHC, van de PasS, Gittenberger-de GrootAC, van ZonneveldAJ, PoelmannRE and HierckBP. (2012). Endothelial colony-forming cells show a mature transcriptional response to shear stress. In Vitro Cell Dev Biol Anim, 48:21–29.
49.
XuJ, LiuX, ChenJ, ZacharekA, CuiX, Savant-BhonsaleS, ChoppM and LiuZ. (2010). Cell-cell interaction promotes rat marrow stromal cell differentiation into endothelial cell via activation of TACE/TNF-alpha signaling. Cell Transplant, 19:43–53.
50.
PlotnikovEY, KhryapenkovaTG, VasilevaAK, MareyMV, GalkinaSI, IsaevNK, ShevalEV, PolyakovVY, SukhikhGT and ZorovDB. (2008). Cell-to-cell cross-talk between mesenchymal stem cells and cardiomyocytes in co-culture. J Cell Mol Med, 12:1622–1631.
51.
CrossMJ and Claesson-WelshL. (2001). FGF and VEGF function in angiogenesis: signalling pathways, biological responses and therapeutic inhibition. Trends Pharmacol Sci, 22:201–207.
52.
OeckinghausA, HaydenMS and GhoshS. (2011). Crosstalk in NF-kappaB signaling pathways. Nat Immunol, 12:695–708.
53.
BakerRG, HaydenMS and GhoshS. (2011). NF-kappaB, inflammation, and metabolic disease. Cell Metab, 13:11–22.
54.
WaniAA, JafarnejadSM, ZhouJ and LiG. (2011). Integrin-linked kinase regulates melanoma angiogenesis by activating NF-kappaB/interleukin-6 signaling pathway. Oncogene, 30:2778–2788.
55.
UchiboriR, TsukaharaT, MizuguchiH, SagaY, UrabeM, MizukamiH, KumeA and OzawaK. (2013). NF-kappaB activity regulates mesenchymal stem cell accumulation at tumor sites. Cancer Res, 73:364–372.
56.
VillarsF, GuillotinB, AmedeeT, DutoyaS, BordenaveL, BareilleR and AmedeeJ. (2002). Effect of HUVEC on human osteoprogenitor cell differentiation needs heterotypic gap junction communication. Am J Physiol Cell Physiol, 282:C775–C785.
57.
KimJ, BreunigMJ, EscalanteLE, BhatiaN, DenuRA, DollarBA, SteinAP, HansonSE, NaderiN, et al. (2012). Biologic and immunomodulatory properties of mesenchymal stromal cells derived from human pancreatic islets. Cytotherapy, 14:925–935.
58.
BartoshTJ, YlostaloJH, BazhanovN, KuhlmanJ and ProckopDJ. (2013). Dynamic compaction of human mesenchymal stem/precursor cells into spheres self-activates caspase-dependent IL1 signaling to enhance secretion of modulators of inflammation and immunity (PGE2, TSG6, and STC1). Stem Cells, 31:2443–2456.
59.
WangL, WaliaB, EvansJ, GewirtzAT, MerlinD and SitaramanSV. (2003). IL-6 induces NF-kappa B activation in the intestinal epithelia. J Immunol, 171:3194–3201.
60.
HanSS, YunH, SonDJ, TompkinsVS, PengL, ChungST, KimJS, ParkES and JanzS. (2010). NF-kappaB/STAT3/PI3K signaling crosstalk in iMyc E mu B lymphoma. Mol Cancer, 9:97
61.
YangJ, LiaoX, AgarwalMK, BarnesL, AuronPE and StarkGR. (2007). Unphosphorylated STAT3 accumulates in response to IL-6 and activates transcription by binding to NFkappaB. Genes Dev, 21:1396–1408.
62.
KochAE, HalloranMM, HaskellCJ, ShahMR and PolveriniPJ. (1995). Angiogenesis mediated by soluble forms of E-selectin and vascular cell adhesion molecule-1. Nature, 376:517–519.
63.
YoshidaT, SinhaS, DandreF, WamhoffBR, HoofnagleMH, KremerBE, WangDZ, OlsonEN and OwensGK. (2003). Myocardin is a key regulator of CArG-dependent transcription of multiple smooth muscle marker genes. Circ Res, 92:856–864.
64.
XieWB, LiZ, MianoJM, LongX and ChenSY. (2011). Smad3-mediated myocardin silencing: a novel mechanism governing the initiation of smooth muscle differentiation. J Biol Chem, 286:15050–15057.
65.
ChenJ, KitchenCM, StrebJW and MianoJM. (2002). Myocardin, a component of a molecular switch for smooth muscle differentiation. J Mol Cell Cardiol, 34:1345–1356.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
