Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
8
pubmed:dateCreated
2011-6-13
pubmed:abstractText
Adult stem cells are important cell sources in regenerative medicine, but isolating them is technically challenging. This study employed a novel strategy to generate stem-like corneal epithelial cells and promote the functional properties of these cells by coculture with embryonic stem cells. The primary corneal epithelial cells were labelled with GFP and cocultured with embryonic stem cells in a transwell or by direct cell-cell contact. The embryonic stem cells were pre-transfected with HSV-tk-puro plasmids and became sensitive to ganciclovir. After 10 days of coculture, the corneal epithelial cells were isolated by treating the cultures with ganciclovir to kill the embryonic stem cells. The expression of stem cell-associated markers (ABCG2, p63) increased whereas the differentiation mark (Keratin 3) decreased in corneal epithelial cells isolated from the cocultures as evaluated by RT-PCR and flow cytometry. Their functional properties of corneal epithelial cells, including cell adhesion, migration and proliferation, were also enhanced. These cells could regenerate a functional stratified corneal epithelial equivalent but did not form tumors. Integrin ?1, phosphorylated focal adhesion kinase and Akt were significantly upregulated in corneal epithelial cells. FAK Inhibitor 14 that suppressed the expression of phosphorylated focal adhesion kinase and Akt inhibited cell adhesion, migration and proliferation. LY294002 that suppressed phosphorylated Akt but not phosphorylated focal adhesion kinase inhibited cell proliferation but had no effect on cell adhesion or migration. These findings demonstrated that the functional properties of stem-like corneal epithelial cells were enhanced by cocultured embryonic stem cells via activation of the integrin ?1-FAK-PI3K/Akt signalling pathway.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
1878-5875
pubmed:author
pubmed:copyrightInfo
Copyright © 2011 Elsevier Ltd. All rights reserved.
pubmed:issnType
Electronic
pubmed:volume
43
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1168-77
pubmed:meshHeading
pubmed-meshheading:21550417-Adult, pubmed-meshheading:21550417-Animals, pubmed-meshheading:21550417-Antigens, CD29, pubmed-meshheading:21550417-Cell Movement, pubmed-meshheading:21550417-Cells, Cultured, pubmed-meshheading:21550417-Coculture Techniques, pubmed-meshheading:21550417-Embryonic Stem Cells, pubmed-meshheading:21550417-Epithelial Cells, pubmed-meshheading:21550417-Focal Adhesion Kinase 2, pubmed-meshheading:21550417-Humans, pubmed-meshheading:21550417-Limbus Corneae, pubmed-meshheading:21550417-Mice, pubmed-meshheading:21550417-Mice, Nude, pubmed-meshheading:21550417-Phosphatidylinositol 3-Kinases, pubmed-meshheading:21550417-Phosphorylation, pubmed-meshheading:21550417-Proto-Oncogene Proteins c-akt, pubmed-meshheading:21550417-Rabbits, pubmed-meshheading:21550417-Signal Transduction, pubmed-meshheading:21550417-Transfection
pubmed:year
2011
pubmed:articleTitle
Enhanced functional properties of corneal epithelial cells by coculture with embryonic stem cells via the integrin ?1-FAK-PI3K/Akt pathway.
pubmed:affiliation
State Key Laboratory of Ophthalmology, Sun yet-sen University, Guangzhou, Guangdong, PR China.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't