We have previously differentiated lung epithelium from human and murine embryonic stem cells (mESCs) and are now exploring the potential applications of these cells, including in the engineering of lung tissue constructs. In this study, we hypothesized that the differentiation and maintenance of lung epithelium derived from mESCs can be enhanced by extracellular matrix (ECM) proteins. Our established differentiation protocol was applied to mESCs grown on a range of ECMs: collagen I, laminin 332, fibronectin, Matrigel, and, as an experimental control, gelatin. The ECMs were coated onto tissue culture plastic (TCP) and poly-DL-lactic acid (PDLLA), a biodegradable polymer we have previously shown to support the growth of mature pneumocytes. Matrigel or Laminin-332 coating of either TCP or PDLLA film resulted in enhanced surfactant protein C gene expression in differentiating mESCs, a direct indication of the upregulation of lung epithelial differentiation. For each combination, changes in the contact angle and ζ potential of protein-coated TCP and PDLLA film confirmed protein adsorption. We conclude that the choice of the coating protein can greatly affect the differentiation of ESCs, and laminin-332-coated PDLLA provided an ECM-degradable scaffold combination that is suitable for engineering of lung tissue constructs.
Get full access to this article
View all access options for this article.
References
1.
RiceW.R., ConkrightJ.J., NaC.L., IkegamiM., ShannonJ.M., WeaverT.E.Maintenance of the mouse type II cell phenotype in vitro. Am J Physiol Lung Cell Mol Physiol, 283:L256. 2002.
2.
AliN.N., EdgarA.J., SamadikuchaksaraeiA., TimsonC.M., RomanskaH.M., PolakJ.M., BishopA.E.Derivation of type II alveolar epithelial cells from murine embryonic stem cells. Tissue Eng, 8:541. 2002.
3.
RipponH.J., AliN.N., PolakJ.M., BishopA.E.Initial observations on the effect of medium composition on the differentiation of murine embryonic stem cells to alveolar type II cells. Cloning Stem Cells, 6:49. 2004.
4.
RipponH.J., PolakJ.M., QinM., BishopA.E.Derivation of distal lung epithelial progenitors from murine embryonic stem cells using a novel three-step differentiation protocol. Stem Cells, 24:1389. 2006.
5.
SamadikuchaksaraeiA., CohenS., IsaacK., RipponH.J., PolakJ.M., BielbyR.C., BishopA.E. Derivation of distal airway epithelium from human embryonic stem cells. Tissue Eng, 12:867. 2006.
Van VrankenB.E., RomanskaH.M., PolakJ.M., RipponH.J., ShannonJ.M., BishopA.E.Coculture of embryonic stem cells with pulmonary mesenchyme: a microenvironment that promotes differentiation of pulmonary epithelium. Tissue Eng, 11:1177. 2005.
8.
AliottaJ.M., PasseroM., MehargJ., KlingerJ., DoonerM.S., PimentelJ., QuesenberryP.J.Stem cells and pulmonary metamorphosis: new concepts in repair and regeneration. J Cell Physiol, 204:725. 2005.
9.
SchugerL., VaraniJ., KillenP.D., SkubitzA.P., GilbrideK.Laminin expression in the mouse lung increases with development and stimulates spontaneous organotypic rearrangement of mixed lung cells. Dev Dyn, 195:43. 1992.
10.
GeppertE.F., WilliamsM.C., MasonR.J.Primary culture of rat alveolar type II Cells on floating collagen membranes. Morphological and biochemical observations. Exp Cell Res, 128:363. 1980.
11.
RannelsS.R., YarnellJ.A., FisherC.S., FabisiakJ.P., RannelsD.E.Role of laminin in maintenance of type II pneumocyte morphology and function. Am J Physiol, 253:C835. 1987.
12.
KawadaH., ShannonJ.M., MasonR.J.Improved maintenance of adult rat alveolar type II cell differentiation in vitro: effect of serum-free, hormonally defined medium and a reconstituted basement membrane. Am J Respir Cell Mol Biol, 3:33. 1990.
13.
ChenP., MarsilioE., GoldsteinR.H., YannasI.V., SpectorM.Formation of lung alveolar-like structures in collagen-glycosaminoglycan scaffolds in vitro. Tissue Eng, 11:1436. 2005.
LevenbergS., GolubJ.S., AmitM., Itskovitz-EldorJ., LangerR.Endothelial cells derived from human embryonic stem cells. Proc Natl Acad Sci USA, 99:4391. 2002.
16.
HutmacherD.W.Scaffolds in tissue engineering bone and cartilage. Biomaterials, 21:2529. 2000.
17.
SafiniaL., BlakerJ.J., MaquetV., BoccacciniA.R., MantalarisA., BismarckA.Characterisation of “wet” polymer surfaces for tissue engineering applications: are flat surfaces a suitable model for complex structures?E-Polymers, 10. 2005.
BakerS.E., DiPasqualeA.P., StockE.L., QuarantaV., FitchmunM., JonesJ.C.Morphogenetic effects of soluble laminin-5 on cultured epithelial cells and tissue explants. Exp Cell Res, 228:262. 1996.
20.
LiS., EdgarD., FasslerR., WadsworthW., YurchencoP.D.The role of laminin in embryonic cell polarization and tissue organization. Dev Cell, 4:613. 2003.
21.
IshiyamaM., TominagaH., ShigaM., SasamotoK., OhkuraY., UenoK.A combined assay of cell viability and in vitro cytotoxicity with a highly water-soluble tetrazolium salt, neutral red and crystal violet. Biol Pharm Bull, 19:1518. 1996.
22.
YasunagaM., TadaS., Torikai-NishikawaS., NakanoY., OkadaM., JaktL.M., NishikawaS., ChibaT., EraT., NishikawaS.Induction and monitoring of definitive and visceral endoderm differentiation of mouse ES cells. Nat Biotechnol, 23:1542. 2005.
23.
KuboA., ShinozakiK., ShannonJ.M., KouskoffV., KennedyM., WooS., FehlingH.J., KellerG.Development of definitive endoderm from embryonic stem cells in culture. Development, 131:1651. 2004.
24.
MfopouJ.K., WillemsE., LeynsL., BouwensL.Expression of regulatory genes for pancreas development during murine embryonic stem cell differentiation. The International journal of developmental biology, 49:915. 2005.
BeersM.F., FisherA.B.Surfactant protein C: a review of its unique properties and metabolism. Am J Physiol, 263:L151. 1992.
27.
BattistaS., GuarnieriD., BorselliC., ZeppetelliS., BorzacchielloA., MayolL., GerbasioD., KeeneD.R., AmbrosioL., NettiP.A.The effect of matrix composition of 3D constructs on embryonic stem cell differentiation. Biomaterials, 26:6194. 2005.
28.
PierceR.A., GriffinG.L., MinerJ.H., SeniorR.M.Expression patterns of laminin alpha1 and alpha5 in human lung during development. Am J Respir Cell Mol Biol, 23:742. 2000.
29.
CorauxC., MeneguzziG., RousselleP., PuchelleE., GaillardD.Distribution of laminin 5, integrin receptors, and branching morphogenesis during human fetal lung development. Dev Dyn, 225:176. 2002.
30.
VirtanenI., LaitinenA., TaniT., PaakkoP., LaitinenL.A., BurgesonR.E., LehtoV.P.Differential expression of laminins and their integrin receptors in developing and adult human lung. Am J Respir Cell Mol Biol, 15:184. 1996.
31.
NguyenN.M., PulkkinenL., SchlueterJ.A., MeneguzziG., UittoJ., SeniorR.M.Lung development in laminin gamma2 deficiency: abnormal tracheal hemidesmosomes with normal branching morphogenesis and epithelial differentiation. Respir Res, 7:28. 2006.
32.
KratochwilK., DziadekM., LohlerJ., HarbersK., JaenischR.Normal epithelial branching morphogenesis in the absence of collagen I. Dev Biol, 117:596. 1986.
33.
RomanJ.Fibronectin and fibronectin receptors in lung development. Exp Lung Res, 23:147. 1997.
34.
AdamsonI.Y., KingG.M., YoungL.Influence of extracellular matrix and collagen components on alveolar type 2 cell morphology and function. In Vitro Cell Dev Biol, 25:494. 1989.
35.
KleinmanH.K., McGarveyM.L., LiottaL.A., RobeyP.G., TryggvasonK., MartinG.R.Isolation and characterization of type IV procollagen, laminin, and heparan sulfate proteoglycan from the EHS sarcoma. Biochemistry, 21:6188. 1982.
36.
SchugerL., O'SheaS., RheinheimerJ., VaraniJ.Laminin in lung development: effects of anti-laminin antibody in murine lung morphogenesis. Dev Biol, 137:26. 1990.
37.
SchugerL., YurchencoP., RelanN.K., YangY.Laminin fragment E4 inhibition studies: basement membrane assembly and embryonic lung epithelial cell polarization requires laminin polymerization. Int J Devel Biol, 42:217. 1998.
38.
MurphyC.L., PolakJ.M.Differentiating embryonic stem cells: GAPDH, but neither HPRT nor beta-tubulin is suitable as an internal standard for measuring RNA levels. Tissue Eng, 8:551. 2002.
39.
NagaharaS., MatsudaT.Cell-substrate and cell-cell interactions differently regulate cytoskeletal and extracellular matrix protein gene expression. J Biomed Mater Res, 32:677. 1996.
40.
BurridgeK., Chrzanowska-WodnickaM.Focal adhesions, contractility, and signaling. Ann Rev Cell Dev Biol, 12:463. 1996.
41.
FarmerS.R., WanK.M., Ben-Ze'evA., PenmanS.Regulation of actin mRNA levels and translation responds to changes in cell configuration. Mol Cell Biol, 3:182. 1983.
42.
OwenG.R., MeredithD.O., ap GwynnI., RichardsR.G.Focal adhesion quantification—a new assay of material biocompatibility?Eur Cells Mater, 9:85. 2005.
43.
ShannonJ.M., PanT., EdeenK.E., NielsenL.D.Influence of the cytoskeleton on surfactant protein gene expression in cultured rat alveolar type II cells. Am J Physiol, 274:L87. 1998.
44.
HeskethJ.E.Sorting of messenger RNAs in the cytoplasm: mRNA localization and the cytoskeleton. Exp Cell Res, 225:219. 1996.
45.
ShannonJ.M., JenningsS.D., NielsenL.D.Modulation of alveolar type II cell differentiated function in vitro. Am J Physiol, 262:L427. 1992.
46.
DeanJ.W., CulbertsonK.C., D'AngeloA.M.Fibronectin and laminin enhance gingival cell attachment to dental implant surfaces in vitro. Int J Oral Maxillofac Implants, 10:721. 1995.
47.
KeslerK.A., HerringM.B., ArnoldM.P., GloverJ.L., ParkH.M., HelmusM.N., BendickP.J.Enhanced strength of endothelial attachment on polyester elastomer and polytetrafluoroethylene graft surfaces with fibronectin substrate. J Vasc Surg, 3:58. 1986.
48.
SeitzT.L., NoonanK.D., HenchL.L., NoonanN.E.Effect of fibronectin on the adhesion of an established cell line to a surface reactive biomaterial. J Biomed Mater Res, 16:195. 1982.
49.
HaynesC.A., SliwinskyE., NordeW.Structural and electrostatic properties of globular-proteins at a polystyrene water interface. J Colloid Interface Sci, 164:394. 1994.
