Mesenchymal stromal cells (MSC) and factors secreted by them are essential components of the hematopoietic stem cell (HSC) niche within the bone marrow microenvironment. It has been shown that the extracellular matrix (ECM) can influence HSC-supportive potential of MSC and is a prerequisite for the proper signaling of morphogens. Therefore, we aimed at the identification of ECM components and candidate morphogens capable of enhancing the expression of HSC-supportive proteins in human MSC, namely, angiopoietin-1 (Ang-1) and stromal cell-derived factor 1 (SDF-1). For this purpose, highly sensitive secreted dual reporter constructs for Ang-1 and SDF-1 were established. These newly designed dual reporter systems enable continuous monitoring of the Ang-1 and SDF-1 promoter activity in an immortalized human MSC line cultured on ECM/morphogen microarrays. Reporter arrays showed that Ang-1 and SDF-1 expression can be induced by different ECM/morphogen combinations. In addition, continuous monitoring of promoter activity allows delineating time-dependent effects of the ECM and morphogens. Thus, we identified that collagen I and vitronectin in combination with Wnt3a favored SDF-1 expression over time, while only transiently inducing the expression of Ang-1. Taken together, the newly developed reporter systems allow for the monitoring of Ang-1 and SDF-1 promoter activity induced by morphogens and the ECM in a combinatorial and high-throughput manner. This technology might therefore be helpful to optimize culture conditions, which favor the activity of MSC as feeder cells for various types of stem and progenitor cells.
Get full access to this article
View all access options for this article.
References
1.
SacchettiB., FunariA., MichienziS., Di CesareS., PiersantiS., SaggioI., TagliaficoE., FerrariS., RobeyP.G., RiminucciM., and BiancoP.Self-renewing osteoprogenitors in bone marrow sinusoids can organize a hematopoietic microenvironment. Cell, 131, 324, 2007.
2.
Mendez-FerrerS., MichurinaT.V., FerraroF., MazloomA.R., MacArthurB.D., LiraS.A., ScaddenD.T., Ma'ayanA., EnikolopovG.N., and FrenetteP.S.Mesenchymal and haematopoietic stem cells form a unique bone marrow niche. Nature, 466, 829, 2010.
3.
SugiyamaT., KoharaH., NodaM., and NagasawaT.Maintenance of the hematopoietic stem cell pool by CXCL12-CXCR4 chemokine signaling in bone marrow stromal cell niches. Immunity, 25, 977, 2006.
4.
SorrentinoA., FerracinM., CastelliG., BiffoniM., TomaselliG., BaiocchiM., FaticaA., NegriniM., PeschleC., and ValtieriM.Isolation and characterization of CD146(+) multipotent mesenchymal stromal cells. Exp Hematol, 36, 1035, 2008.
IchiiM., FrankM.B., IozzoR.V., and KincadeP.W.The canonical Wnt pathway shapes niches supportive of hematopoietic stem/progenitor cells. Blood, 119, 1683, 2012.
10.
IkushimaY.M., AraiF., HosokawaK., ToyamaH., TakuboK., FuruyashikiT., NarumiyaS., and SudaT.Prostaglandin E2 regulates murine hematopoietic stem/progenitor cells directly via EP4 receptor and indirectly through mesenchymal progenitor cells. Blood, 121, 1995, 2013.
11.
SeiffertD.Detection of vitronectin in mineralized bone matrix. J Histochem Cytochem, 44, 275, 1996.
12.
NilssonS.K., DebatisM.E., DoonerM.S., MadriJ.A., QuesenberryP.J., and BeckerP.S.Immunofluorescence characterization of key extracellular matrix proteins in murine bone marrow in situ. J Histochem Cytochem, 46, 371, 1998.
13.
NilssonS.K., JohnstonH.M., WhittyG.A., WilliamsB., WebbR.J., DenhardtD.T., BertoncelloI., BendallL.J., SimmonsP.J., and HaylockD.N.Osteopontin, a key component of the hematopoietic stem cell niche and regulator of primitive hematopoietic progenitor cells. Blood, 106, 1232, 2005.
14.
StierS., KoY., ForkertF., LutzC., NeuhausT., GrunewaldE., ChengT., DombkowskiD., CalviL.M., RittlingS.R., and ScaddenD.T.Osteopontin is a hematopoietic stem cell niche component that negatively regulates stem cell pool size. J Exp Med, 201, 1781, 2005.
15.
DochevaD., PopovC., MutschlerW., and SchiekerM.Human mesenchymal stem cells in contact with their environment: surface characteristics and the integrin system. J Cell Mol Med, 11, 21, 2007.
16.
SeibF.P., MüllerK., FrankeM., GrimmerM., BornhäuserM., and WernerC.Engineered extracellular matrices modulate the expression profile and feeder properties of bone marrow-derived human multipotent mesenchymal stromal cells. Tissue Eng Part A, 15, 3161, 2009.
17.
LanferB., SeibF.P., FreudenbergU., StamovD., BleyT., BornhäuserM., and WernerC.The growth and differentiation of mesenchymal stem and progenitor cells cultured on aligned collagen matrices. Biomaterials, 30, 5950, 2009.
18.
HuangN.F., ChuJ., LeeR.J., and LiS.Biophysical and chemical effects of fibrin on mesenchymal stromal cell gene expression. Acta Biomater, 6, 3947, 2010.
19.
LaiY.L., SunY., SkinnerC.M., SonE.L., LuZ.D., TuanR.S., JilkaR.L., LingJ.A., and ChenX.D.Reconstitution of marrow-derived extracellular matrix ex vivo: a robust culture system for expanding large-scale highly functional human mesenchymal stem cells. Stem Cells Dev, 19, 1095, 2010.
20.
PeiM., HeF., and KishV.L.Expansion on extracellular matrix deposited by human bone marrow stromal cells facilitates stem cell proliferation and tissue-specific lineage potential. Tissue Eng Part A, 17, 3067, 2011.
21.
PrewitzM.C., SeibF.P., von BoninM., FriedrichsJ., StisselA., NiehageC., MüllerK, AnastassiadisK., WaskowC., HoflackB., BornhäuserM., and WernerC.Tightly anchored tissue-mimetic matrices as instructive stem cell microenvironments. Nat Methods, 10, 788, 2013.
22.
KleinG.The Extracellular-matrix of the hematopoietic microenvironment. Experientia, 51, 914, 1995.
23.
LinF.B., RenX.D., PanZ., MacriL., ZongW.X., TonnesenM.G., RafailovichM., Bar-SagiD., and ClarkR.A.F.Fibronectin growth factor-binding domains are required for fibroblast survival. J Invest Dermatol, 131, 84, 2011.
24.
WijelathE.S., RahmanS., NamekataM., MurrayJ., NishimuraT., Mostafavi-PourZ., PatelY., SudaY., HumphriesM.J., and SobelM.Heparin-II domain of fibronectin is a vascular endothelial growth factor-binding domain—enhancement of VEGF biological activity by a singular growth factor/matrix protein synergism. Circ Res, 99, 853, 2006.
25.
RahmanS., PatelY., MurrayJ., PatelK.V., SumathipalaR., SobelM., and WijelathE.S.Novel hepatocyte growth factor (HGF) binding domains on fibronectin and vitronectin coordinate a distinct and amplified Met-integrin induced signalling pathway in endothelial cells. BMC Cell Biol, 6, 8, 2005.
26.
BockerW., YinZ., DrosseI., HaastersF., RossmannO., WiererM., PopovC., LocherM., MutschlerW., DochevaD., and SchiekerM.Introducing a single-cell-derived human mesenchymal stem cell line expressing hTERT after lentiviral gene transfer. J Cell Mol Med, 12, 1347, 2008
27.
ZuffereyR., DullT., MandelR.J., BukovskyA., QuirozD., NaldiniL., and TronoD.Self-inactivating lentivirus vector for safe and efficient in vivo gene delivery. J Virol, 72, 9873, 1998.
28.
SchambachA., BohneJ., ChandraS., WillE., MargisonG.P., WilliamsD.A., and BaumC.Equal potency of gammaretroviral and lentiviral SIN vectors for expression of O-6-methylguanine-DNA methyltransferase in hematopoietic cells. Mol Ther, 13, 391, 2006.
29.
ReyaT., O'RiordanM., OkamuraR., DevaneyE., WillertK., NusseR., and GrosschedlR.Wnt signaling regulates B lymphocyte proliferation through a LEF-1 dependent mechanism. Immunity, 13, 15, 2000.
30.
HosokawaK., AraiF., YoshiharaH., IwasakiH., HembreeM., YinT., NakamuraY., GomeiY., TakuboK., ShiamaH., MatsuokaS., LiL., and SudaT.Cadherin-based adhesion is a potential target for niche manipulation to protect hematopoietic stem cells in adult bone marrow. Cell Stem Cell, 6, 194, 2010.
31.
DishowitzM.I., ZhuF., SundararaghavanH.G., IfkovitsJ.L., BurdickJ.A., and HankensonK.D.Jagged1 immobilization to an osteoconductive polymer activates the Notch signaling pathway and induces osteogenesis. J Biomed Mater Res A, 2013 [Epub ahead of print]; DOI: 10.1002/jbm.a.34825.
32.
GobaaS., HoehnelS., RoccioM., NegroA., KobelS., and LutolfM.P.Artificial niche microarrays for probing single stem cell fate in high throughput. Nat Methods, 8, 949, 2011.
33.
PeeraniR., and ZandstraP.W.Enabling stem cell therapies through synthetic stem cell-niche engineering. J Clin Invest, 120, 60, 2010.
34.
AndersonD.G., LevenbergS., and LangerR.Nanoliter-scale synthesis of arrayed biomaterials and application to human embryonic stem cells. Nat Biotechnol, 22, 863, 2004.
35.
FlaimC.J., ChienS., and BhatiaS.N.An extracellular matrix microarray for probing cellular differentiation. Nat Methods, 2, 119, 2005.
36.
SoenY., MoriA., PalmerT.D., and BrownP.O.Exploring the regulation of human neural precursor cell differentiation using arrays of signaling microenvironments. Mol Syst Biol, 2, 37, 2006.
37.
LutolfM.P., DoyonnasR., HavenstriteK., KoleckarK., and BlauH.M.Perturbation of single hematopoietic stem cell fates in artificial niches. Integr Biol (Camb), 1, 59, 2009.
38.
BergerJ., HauberJ., HauberR., GeigerR., and CullenB.R.Secreted placental alkaline-phosphatase—a powerful new quantitative indicator of gene-expression in eukaryotic cells. Gene, 66, 1, 1988.
39.
TannousB.A.Gaussia luciferase reporter assay for monitoring biological processes in culture and in vivo. Nat Protocols, 4, 582, 2009.
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.
