title: Comparative proteomic analysis of supportive and unsupportive extracellular matrix substrates for human embryonic stem cell maintenance.
abstract: Human embryonic stem cells (hESCs) are pluripotent cells that have indefinite replicative potential and the ability to differentiate into derivatives of all three germ layers. hESCs are conventionally grown on mitotically inactivated mouse embryonic fibroblasts (MEFs) or feeder cells of human origin. In addition, feeder-free culture systems can be used to support hESCs, in which the adhesive substrate plays a key role in the regulation of stem cell self-renewal or differentiation. Extracellular matrix (ECM) components define the microenvironment of the niche for many types of stem cells, but their role in the maintenance of hESCs remains poorly understood. We used a proteomic approach to characterize in detail the composition and interaction networks of ECMs that support the growth of self-renewing hESCs. Whereas many ECM components were produced by supportive and unsupportive MEF and human placental stromal fibroblast feeder cells, some proteins were only expressed in supportive ECM, suggestive of a role in the maintenance of pluripotency. We show that identified candidate molecules can support attachment and self-renewal of hESCs alone (fibrillin-1) or in combination with fibronectin (perlecan, fibulin-2), in the absence of feeder cells. Together, these data highlight the importance of specific ECM interactions in the regulation of hESC phenotype and provide a resource for future studies of hESC self-renewal.
authors: North West Embryonic Stem Cell Centre, Faculty of Life Sciences, University of Manchester, Manchester M13 9NT, United Kingdom.
authors: Despina
authors: D
authors: Soteriou
authors: Banu
authors: B
authors: Iskender
authors: Adam
authors: A
authors: Byron
authors: Jonathan D
authors: JD
authors: Humphries
authors: Simon
authors: S
authors: Borg-Bartolo
authors: Marie-Claire
authors: MC
authors: Haddock
authors: Melissa A
authors: MA
authors: Baxter
authors: David
authors: D
authors: Knight
authors: Martin J
authors: MJ
authors: Humphries
authors: Susan J
authors: SJ
authors: Kimber
mesh_terms: D000818:Animals; D002135:Calcium-Binding Proteins / Q000378:metabolism; D018929:Cell Culture Techniques; D016000:Cluster Analysis; D017077:Culture Media, Conditioned / Q000737:chemistry; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism*; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D061252:Feeder Cells; D000071838:Fibrillin-1; D000071837:Fibrillins; D005347:Fibroblasts / Q000166:cytology; D005353:Fibronectins / Q000378:metabolism; D019812:Heparan Sulfate Proteoglycans / Q000378:metabolism; D006801:Humans; D016023:Integrins / Q000378:metabolism; D007621:Karyotyping; D051379:Mice; D008840:Microfilament Proteins / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D054392:Adipokines
chemical_list: D002135:Calcium-Binding Proteins; D017077:Culture Media, Conditioned; D016326:Extracellular Matrix Proteins; D000071838:Fibrillin-1; D000071837:Fibrillins; D005353:Fibronectins; D019812:Heparan Sulfate Proteoglycans; D016023:Integrins; D008840:Microfilament Proteins; C000605722:FBN1 protein, human; D054392:Adipokines; C000605724:Fbn1 protein, mouse; C084612:fibulin 2; C071927:perlecan
keywords: Cell Biology; Embryonic Stem Cell; Extracellular Matrix; Extracellular Matrix Proteins; Proteomics; Stem Cells
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