Engineered Extracellular Matrices Modulate the Expression Profile and Feeder Properties of Bone Marrow-Derived Human Multipotent Mesenchymal Stromal Cells
Restricted accessResearch articleFirst published online October, 2009
Engineered Extracellular Matrices Modulate the Expression Profile and Feeder Properties of Bone Marrow-Derived Human Multipotent Mesenchymal Stromal Cells
The bone marrow harbors multipotent mesenchymal stromal cells (MSCs) that nurture hematopoietic stem cells (HSCs). The extracellular matrix (ECM) is an integral part of the bone marrow, and the aim of this study was therefore to examine the effect of engineered ECM substrates on MSC gene expression over time and to determine quantitatively the functional ability of ECM-cultured MSCs to support HSCs. ECMs were surface immobilized using thin films of maleic anhydride to covalently immobilize tropocollagen or fibrillar collagen type I to the substrate. Where indicated, collagen type I fibrils were supplemented with heparin or hyaluronic acid. All surfaces maintained MSC viability and supported cell expansion. Microarray analysis of MSCs cultured on engineered ECM substrates revealed that culture time, as well as substrate composition, significantly affected expression levels. Based on these studies, it was possible to predict the effect of these substrates on in vitro HSC clonogenicity and self-renewal. The ability to regulate the expression of stromal factors using engineered ECM is exciting and warrants further studies to identify the ECM components and combinations that maximize the expansion of clonogenic HSCs.
LichtmanM.A.The ultrastructure of the hemopoietic environment of the marrow: a review. Exp Hematol, 9:391. 1981.
4.
Lo CelsoC., FlemingH.E., WuJ.W., ZhaoC.X., Miake-LyeS., FujisakiJ., CoteD., RoweD.W., LinC.P., ScaddenD.T.Live-animal tracking of individual haematopoietic stem/progenitor cells in their niche. Nature, 457:92. 2009.
5.
DaleyW.P., PetersS.B., LarsenM.Extracellular matrix dynamics in development and regenerative medicine. J Cell Sci, 121:255. 2008.
6.
FriedensteinA.J.HeerscheJ N. M., KanisJ. A.Osteogenic stem cells in the bone marrow. Bone and Mineral Research. Amsterdam: Elsevier Science Publishers (Biomedical Division), 1990; 243–272.
7.
BanfiA., MuragliaA., DozinB., MastrogiacomoM., CanceddaR., QuartoR.Proliferation kinetics and differentiation potential of ex vivo expanded human bone marrow stromal cells: implications for their use in cell therapy. Exp Hematol, 28:707. 2000.
8.
BaxterM.A., WynnR.F., JowittS.N., WraithJ.E., FairbairnL.J., BellantuonoI.Study of telomere length reveals rapid aging of human marrow stromal cells following in vitro expansion. Stem Cells, 22:675. 2004.
9.
DigirolamoC.M., StokesD., ColterD., PhinneyD.G., ClassR., ProckopD.J.Propagation and senescence of human marrow stromal cells in culture: a simple colony-forming assay identifies samples with the greatest potential to propagate and differentiate. Br J Haematol, 107:275. 1999.
10.
SeibF.P., FrankeM., JingD., WernerC., BornhauserM.Endogenous bone morphogenetic proteins in human bone marrow-derived multipotent mesenchymal stromal cells. Eur J Cell Biol, 88:257. 2009.
11.
WagnerW., HornP., CastoldiM., DiehlmannA., BorkS., SaffrichR., BenesV., BlakeJ., PfisterS., EcksteinV., HoA.D.Replicative senescence of mesenchymal stem cells: a continuous and organized process. PLoS ONE, 3:e2213. 2008.
12.
ChenX.D., DusevichV., FengJ.Q., ManolagasS.C., JilkaR.L.Extracellular matrix made by bone marrow cells facilitates expansion of marrow-derived mesenchymal progenitor cells and prevents their differentiation into osteoblasts. J Bone Miner Res, 22:1943. 2007.
13.
MatsubaraT., TsutsumiS., PanH., HiraokaH., OdaR., NishimuraM., KawaguchiH., NakamuraK., KatoY.A new technique to expand human mesenchymal stem cells using basement membrane extracellular matrix. Biochem Biophys Res Commun, 313:503. 2004.
14.
MauneyJ.R., Kirker-HeadC., AbrahamsonL., GronowiczG., VollochV., KaplanD.L.Matrix-mediated retention of in vitro osteogenic differentiation potential and in vivo bone-forming capacity by human adult bone marrow-derived mesenchymal stem cells during ex vivo expansion. J Biomed Mater Res A, 79:464. 2006.
15.
MauneyJ.R., VollochV., KaplanD.L.Matrix-mediated retention of adipogenic differentiation potential by human adult bone marrow-derived mesenchymal stem cells during ex vivo expansion. Biomaterials, 26:6167. 2005.
16.
DattaN., HoltorfH.L., SikavitsasV.I., JansenJ.A., MikosA.G.Effect of bone extracellular matrix synthesized in vitro on the osteoblastic differentiation of marrow stromal cells. Biomaterials, 26:971. 2005.
17.
KleesR.F., SalasznykR.M., KingsleyK., WilliamsW.A., BoskeyA., PlopperG.E.Laminin-5 induces osteogenic gene expression in human mesenchymal stem cells through an ERK-dependent pathway. Mol Biol Cell, 16:881. 2005.
18.
SalasznykR.M., KleesR.F., WilliamsW.A., BoskeyA., PlopperG.E.Focal adhesion kinase signaling pathways regulate the osteogenic differentiation of human mesenchymal stem cells. Exp Cell Res, 313:22. 2007.
SalchertK., OswaldJ., StrellerU., GrimmerM., HeroldN., WernerC.Fibrillar collagen assembled in the presence of glycosaminoglycans to constitute bioartificial stem cell niches in vitro. J Mater Sci Mater Med, 16:581. 2005.
21.
SalchertK., StrellerU., PompeT., HeroldN., GrimmerM., WernerC.In vitro reconstitution of fibrillar collagen type I assemblies at reactive polymer surfaces. Biomacromolecules, 5:1340. 2004.
22.
SalchertK., PompeT., SperlingC., WernerC.Quantitative analysis of immobilized proteins and protein mixtures by amino acid analysis. J Chromatogr A, 1005:113. 2003.
23.
OswaldJ., BoxbergerS., JorgensenB., FeldmannS., EhningerG., BornhauserM., WernerC.Mesenchymal stem cells can be differentiated into endothelial cells in vitro. Stem Cells, 22:377. 2004.
24.
OswaldJ., SteudelC., SalchertK., JoergensenB., ThiedeC., EhningerG., WernerC., BornhauserM.Gene-expression profiling of CD34+ hematopoietic cells expanded in a collagen I matrix. Stem Cells, 24:494. 2006.
25.
PloemacherR.E.FreshneyR. I., PragnellI. B., FreshneyM. G.Cobblestone area forming cell (CACF) assay. Culture of Hematopoietic Cells. New York: Wiley-Liss, 1994; 1–21.
26.
KanayaK., OkayamaS.Penetration and energy-loss theory of electrons in solid targets. J Phys D Appl Phys, 5:43. 1972.
27.
QiH., AguiarD.J., WilliamsS.M., La PeanA., PanW., VerfaillieC.M.Identification of genes responsible for osteoblast differentiation from human mesodermal progenitor cells. Proc Natl Acad Sci U S A, 100:3305. 2003.
28.
DoiM., NaganoA., NakamuraY.Molecular cloning and characterization of a novel gene, EMILIN-5, and its possible involvement in skeletal development. Biochem Biophys Res Commun, 313:888. 2004.
29.
DjouadF., BonyC., HauplT., UzeG., LahlouN., Louis-PlenceP., ApparaillyF., CanovasF., RemeT., SanyJ., JorgensenC., NoelD.Transcriptional profiles discriminate bone marrow-derived and synovium-derived mesenchymal stem cells. Arthritis Res Ther, 7:R1304. 2005.
30.
JeongJ.A., HongS.H., GangE.J., AhnC., HwangS.H., YangI.H., HanH., KimH.Differential gene expression profiling of human umbilical cord blood-derived mesenchymal stem cells by DNA microarray. Stem Cells, 23:584. 2005.
31.
WagnerW., WeinF., SeckingerA., FrankhauserM., WirknerU., KrauseU., BlakeJ., SchwagerC., EcksteinV., AnsorgeW., HoA.D.Comparative characteristics of mesenchymal stem cells from human bone marrow, adipose tissue, and umbilical cord blood. Exp Hematol, 33:1402. 2005.
32.
SongL., WebbN.E., SongY., TuanR.S.Identification and functional analysis of candidate genes regulating mesenchymal stem cell self-renewal and multipotency. Stem Cells, 24:1707. 2006.
33.
DjouadF., DelormeB., MauriceM., BonyC., ApparaillyF., Louis-PlenceP., CanovasF., CharbordP., NoelD., JorgensenC.Microenvironmental changes during differentiation of mesenchymal stem cells towards chondrocytes. Arthritis Res Ther, 9:R33. 2007.
34.
KultererB., FriedlG., JandrositzA., Sanchez-CaboF., ProkeschA., PaarC., ScheidelerM., WindhagerR., PreiseggerK.H., TrajanoskiZ.Gene expression profiling of human mesenchymal stem cells derived from bone marrow during expansion and osteoblast differentiation. BMC Genomics, 8:70. 2007.
35.
OhnishiS., YasudaT., KitamuraS., NagayaN.Effect of hypoxia on gene expression of bone marrow-derived mesenchymal stem cells and mononuclear cells. Stem Cells, 25:1166. 2007.
36.
PedemonteE., BenvenutoF., CasazzaS., MancardiG., OksenbergJ.R., UccelliA., BaranziniS.E.The molecular signature of therapeutic mesenchymal stem cells exposes the architecture of the hematopoietic stem cell niche synapse. BMC Genomics, 8:65. 2007.
37.
TsaiM.S., HwangS.M., ChenK.D., LeeY.S., HsuL.W., ChangY.J., WangC.N., PengH.H., ChangY.L., ChaoA.S., ChangS.D., LeeK.D., WangT.H., WangH.S., SoongY.K.Functional network analysis of the transcriptomes of mesenchymal stem cells derived from amniotic fluid, amniotic membrane, cord blood, and bone marrow. Stem Cells, 25:2511. 2007.
38.
CovasD.T., PanepucciR.A., FontesA.M., SilvaW.A.Jr., OrellanaM.D., FreitasM.C., NederL., SantosA.R., PeresL.C., JamurM.C., ZagoM.A.Multipotent mesenchymal stromal cells obtained from diverse human tissues share functional properties and gene-expression profile with CD146+ perivascular cells and fibroblasts. Exp Hematol, 36:642. 2008.
39.
TondreauT., DejeneffeM., MeulemanN., StamatopoulosB., DelforgeA., MartiatP., BronD., LagneauxL.Gene expression pattern of functional neuronal cells derived from human bone marrow mesenchymal stromal cells. BMC Genomics, 9:166. 2008.
40.
CampbellA.D., WichaM.S.Extracellular matrix and the hematopoietic microenvironment. J Lab Clin Med, 112:140. 1988.
41.
GordonM.Y.Extracellular matrix of the marrow microenvironment. Br J Haematol, 70:1. 1988.
42.
HaylockD.N., NilssonS.K.The role of hyaluronic acid in hemopoietic stem cell biology. Regen Med, 1:437. 2006.
43.
HackneyJ.A., CharbordP., BrunkB.P., StoeckertC.J., LemischkaI.R., MooreK.A.A molecular profile of a hematopoietic stem cell niche. Proc Natl Acad Sci U S A, 99:13061. 2002.
44.
CharbordP., MooreK.Gene expression in stem cell-supporting stromal cell lines. Ann N Y Acad Sci, 1044:159. 2005.
45.
PeledA., PetitI., KolletO., MagidM., PonomaryovT., BykT., NaglerA., Ben-HurH., ManyA., ShultzL., LiderO., AlonR., ZiporiD., LapidotT.Dependence of human stem cell engraftment and repopulation of NOD/SCID mice on CXCR4. Science, 283:845. 1999.
46.
NieY., HanY.C., ZouY.R.CXCR4 is required for the quiescence of primitive hematopoietic cells. J Exp Med, 205:777. 2008.
47.
AraiF., HiraoA., OhmuraM., SatoH., MatsuokaS., TakuboK., ItoK., KohG.Y., SudaT.Tie2/angiopoietin-1 signaling regulates hematopoietic stem cell quiescence in the bone marrow niche. Cell, 118:149. 2004.
48.
SacchettiB., FunariA., MichienziS., Di CesareS., PiersantiS., SaggioI., TagliaficoE., FerrariS., RobeyP.G., RiminucciM., BiancoP.Self-renewing osteoprogenitors in bone marrow sinusoids can organize a hematopoietic microenvironment. Cell, 131:324. 2007.
BreemsD.A., BloklandE.A., NebenS., PloemacherR.E.Frequency analysis of human primitive haematopoietic stem cell subsets using a cobblestone area forming cell assay. Leukemia, 8:1095. 1994.
51.
SutherlandH.J., LansdorpP.M., HenkelmanD.H., EavesA.C., EavesC.J.Functional characterization of individual human hematopoietic stem cells cultured at limiting dilution on supportive marrow stromal layers. Proc Natl Acad Sci U S A, 87:3584. 1990.
52.
CampbellA., WichaM.S., LongM.Extracellular matrix promotes the growth and differentiation of murine hematopoietic cells in vitro. J Clin Invest, 75:2085. 1985.
53.
MergenthalerH.G., DormerP.Hemopoiesis in human micro long-term bone marrow culture with preformed extracellular matrix. Haematologica, 75:12. 1990.
54.
TeofiliL., SargiacomoM., IovinoM.S., ZiniG., LeoneG., BizziB., PeschleC.Effects of a preformed extracellular matrix on long-term serum-free bone marrow culture. Ann Hematol, 65:22. 1992.
55.
CampbellA.D., LongM.W., WichaM.S.Haemonectin, a bone marrow adhesion protein specific for cells of granulocyte lineage. Nature, 329:744. 1987.
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.
