Biomaterial matrices presenting extracellular matrix (ECM) components in a controlled three-dimensional configuration provide a unique system to study neural stem cell (NSC)–ECM interactions. We cultured primary murine neurospheres in a methylcellulose (MC) scaffold functionalized with laminin-1 (MC-x-LN1) and monitored NSC survival, apoptosis, migration, differentiation, and matrix production. Overall, MC-x-LN1 enhanced both NSC survival and maturation compared with MC controls. Significantly lower levels of apoptotic activity were observed in MC-x-LN1 than in MC controls, as measured by bcl-2/bax gene expression and tetramethylrhodamine-dUTP nick end labeling. A higher percentage of NSCs extended neurites in a β1-integrin-mediated fashion in MC-x-LN1 than in MC controls. Further, the differentiation profiles of NSCs in MC-x-LN1 exhibited higher levels of neuronal and oligodendrocyte precursor markers than in MC controls. LN1 production and co-localization with α6β1 integrins was markedly increased within MC-x-LN1, whereas the production of fibronectin was more pronounced in MC controls. These findings demonstrate that NSC microenvironments modulate cellular activity throughout the neurosphere, contributing to our understanding of ECM-mediated NSC behavior and provide new avenues for developing rationally designed couriers for neurotransplantation.
Get full access to this article
View all access options for this article.
References
1.
LiS., HarrisonD., CarbonettoS., FasslerR., SmythN., EdgarD., YurchencoP.D.Matrix assembly, regulation, and survival functions of laminin and its receptors in embryonic stem cell differentiation. J Cell Biol, 157:1279. 2002.
LeoneD.P., RelvasJ.B., CamposL.S., HemmiS., BrakebuschC., FassleR., Ffrench-ConstantC., SuterU. Regulation of neural progenitor proliferation and survival by b1 integrins. J Cell Sci, 118:2589. 2005.
4.
TateM.C., GarciaA.J., KeselowskyB.G., SchummM.A., ArcherD.R., LaPlacaM.C.Specific beta-1 integrins mediate adhesion, migration, and differentiation of neural progenitors derived from the embryonic striatum. Mol Cell Neurosci, 27:22. 2004.
5.
JacquesT.S., RelvasJ.B., NishimuraS., PytelaR., EdwardsG.M., StreuliC.H., Ffrench-ConstantC.Neural precursor cell chain migration and division are regulated through different beta 1 integrins. Development, 125:3167. 1998.
6.
CamposL.S., LeoneD.P., RelvasJ.B., BrakebuschC., FasslerR., SuterU., Ffrench-ConstantC.Beta1 integrins activate a MAPK signalling pathway in neural stem cells that contributes to their maintenance. Development, 131:3433. 2004.
7.
KereverA., SchnackJ., VellingaD., IchikawaN., MoonC., Arikawa-HirasawaE., EfirdJ.T., MercierF.Novel extracellular matrix structures in the neural stem cell niche capture the neurogenic factor FGF-2 from the extracellular milieu. Stem Cells, 25:2146. 2007.
8.
MahoneyM.J., AnsethK.S.Contrasting effects of collagen and bFGF-2 on neural cell function in degradable synthetic PEG hydrogels. J Biomed Mater Res A, 81:269. 2007.
9.
SahaK., IrwinE.F., KozhukhJ., SchafferD.V., HealyK.E.Biomimetic interfacial interpenetrating polymer networks control neural stem cell behavior. J Biomed Mater Res A, 81:240. 2007.
10.
NakajimaM., IshimuroT., KatoK., KoI.K., HirataI., ArimaY., IwataH.Combinatorial protein display for the cell-based screening of biomaterials that direct neural stem cell differentiation. Biomaterials, 28:1040. 2007.
11.
CukiermanE., PankovR., StevensD.R., YamadaK.M.Taking cell-matrix adhesions to the third dimension. Science, 294:1708. 2001.
12.
FriedlP., BrockerE.B.The biology of cell locomotion within three-dimensional extracellular matrix. Cell Mol Life Sci, 57:41. 2000.
13.
TateM.C., ShearD.A., HoffmanS.W., SteinD.G., LaPlacaM.C.Biocompatibility of methylcellulose-based constructs designed for intracerebral gelation following experimental traumatic brain injury. Biomaterials, 22:1113. 2001.
14.
StabenfeldtS.E., GarciaA.J., LaPlacaM.C.Thermoreversible laminin-functionalized hydrogel for neural tissue engineering. J Biomed Mater Res A, 77A:718. 2006.
15.
GuptaD., TatorC.H., ShoichetM.S.Fast-gelling injectable blend of hyaluronan and methylcellulose for intrathecal, localized delivery to the injured spinal cord. Biomaterials, 27:2370. 2006.
16.
WellsM.R., KrausK., BatterD.K., BluntD.G., WeremowitzJ., LynchS.E., AntoniadesH.N., HanssonH.A.Gel matrix vehicles for growth factor application in nerve gap injuries repaired with tubes: a comparison of biomatrix, collagen, and methylcellulose. Exp Neurol, 146:395. 1997.
17.
BakshiA., KeckC.A., KoshkinV.S., LeBoldD.G., SimanR., SnyderE.Y., McIntoshT.K.Caspase-mediated cell death predominates following engraftment of neural progenitor cells into traumatically injured rat brain. Brain Res, 1065:8. 2005.
18.
MarchioniniD.M., CollierT.J., CamargoM., McGuireS., PitzerM., SortwellC.E.Interference with anoikis-induced cell death of dopamine neurons: implications for augmenting embryonic graft survival in a rat model of Parkinson's disease. J Comp Neurol, 464:172. 2003.
19.
TateM.C., ShearD.A., HoffmanS.W., SteinD.G., ArcherD.R., LaPlacaM.C.Fibronectin promotes survival and migration of primary neural stem cells transplanted into the traumatically injured mouse brain. Cell Transplant, 11:283. 2002.
20.
PinkseG.G., BouwmanW.P., Jiawan-LalaiR., TerpstraO.T., BruijnJ.A., de HeerE.Integrin signaling via RGD peptides and anti-beta1 antibodies confers resistance to apoptosis in islets of Langerhans. Diabetes, 55:312. 2006.
21.
TateC.C., ShearD.A., TateM.C., ArcherD.R., SteinD.G., LaPlacaM.C.Laminin and fibronectin scaffolds enhance neural stem cell transplantation into the injured brain. J Tissue Eng Regen Med, 3:208. 2009.
22.
OkabeS., ForsbergNilssonK., SpiroA.C., SegalM., McKayR.D.G.Development of neuronal precursor cells and functional postmitotic neurons from embryonic stem cells in vitro. Mech Dev, 59:89. 1996.
DeckerT., Lohmann-MatthesM.L.A quick and simple method for the quantitation of lactate dehydrogenase release in measurements of cellular cytotoxicity and tumor necrosis factor (TNF) activity. J Immunol Methods, 115:61. 1988.
25.
WatanabeK., NakamuraM., OkanoH., ToyamaY.Establishment of three-dimensional culture of neural stem/progenitor cells in collagen Type-1 Gel. Restor Neurol Neurosci, 25:109. 2007.
26.
ByersB.A., PavlathG.K., MurphyT.J., KarsentyG., GarciaA.J.Cell-type-dependent up-regulation of in vitro mineralization after overexpression of the osteoblast-specific transcription factor Runx2/Cbfa1. J Bone Miner Res, 17:1931. 2002.
27.
BoleyS.E., WongV.A., FrenchJ.E., RecioL.p53 heterozygosity alters the mRNA expression of p53 target genes in the bone marrow in response to inhaled benzene. Toxicol Sci, 66:209. 2002.
28.
BrazelC.Y., LimkeT.L., OsborneJ.K., MiuraT., CaiJ., PevnyL., RaoM.S.Sox2 expression defines a heterogeneous population of neurosphere-forming cells in the adult murine brain. Aging Cell, 4:197. 2005.
29.
WilliamsC.M., EnglerA.J., SloneR.D., GalanteL.L., SchwarzbauerJ.E.Fibronectin expression modulates mammary epithelial cell proliferation during acinar differentiation. Cancer Res, 68:3185. 2008.
30.
ChaoD.T., KorsmeyerS.J.BCL-2 family: regulators of cell death. Annu Rev Immunol, 16:395. 1998.
31.
HallP.E., LathiaJ.D., MillerN.G., CaldwellM.A., Ffrench-ConstantC.Integrins are markers of human neural stem cells. Stem Cells, 24:2078. 2006.
32.
MinerJ.H., LiC., MuddJ.L., GoG., SutherlandA.E.Compositional and structural requirements for laminin and basement membranes during mouse embryo implantation and gastrulation. Development, 131:2247. 2004.
33.
PinkseG.G., VoorhoeveM.P., NotebornM., TerpstraO.T., BruijnJ.A., De HeerE.Hepatocyte survival depends on beta1-integrin-mediated attachment of hepatocytes to hepatic extracellular matrix. Liver Int, 24:218. 2004.
34.
MolcanyiM., RiessP., BentzK., MaegeleM., HeschelerJ., SchafkeB., TrappT., NeugebauerE., KlugN., SchaferU.Trauma-associated inflammatory response impairs embryonic stem cell survival and integration after implantation into injured rat brain. J Neurotrauma, 24:625. 2007.
LiesiP., FriedG., StewartR.R.Neurons and glial cells of the embryonic human brain and spinal cord express multiple and distinct isoforms of laminin. J Neurosci Res, 64:144. 2001.
39.
HiraokaM., KatoK., Nakaji-HirabayashiT., IwataH.Enhanced survival of neural cells embedded in hydrogels composed of collagen and laminin-derived cell adhesive peptide. Bioconjug Chem, 20:976. 2009.
40.
SilvaG.A., CzeislerC., NieceK.L., BeniashE., HarringtonD.A., KesslerJ.A., StuppS.I.Selective differentiation of neural progenitor cells by high-epitope density nanofibers. Science, 303:1352. 2004.
BanerjeeA., ArhaM., ChoudharyS., AshtonR.S., BhatiaS.R., SchafferD.V., KaneR.S.The influence of hydrogel modulus on the proliferation and differentiation of encapsulated neural stem cells. Biomaterials, 30:4695. 2009.
43.
HynesS.R., RauchM.F., BertramJ.P., LavikE.B.A library of tunable poly(ethylene glycol)/poly(L-lysine) hydrogels to investigate the material cues that influence neural stem cell differentiation. J Biomed Mater Res A, 89A:499. 2009.
44.
LeipzigN.D., XuC., ZahirT., ShoichetM.S.Functional immobilization of interferon-gamma induces neuronal differentiation of neural stem cells. J Biomed Mater Res A, 93:625. 2010.
45.
SeidlitsS.K., KhaingZ.Z., PetersenR.R., NickelsJ.D., VanscoyJ.E., ShearJ.B., SchmidtC.E.The effects of hyaluronic acid hydrogels with tunable mechanical properties on neural progenitor cell differentiation. Biomaterials, 31:3930. 2010.
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.
