@article {1363130,
	title = {TGF-β3 stimulates stromal matrix assembly by human corneal keratocyte-like cells},
	journal = {Invest Ophthalmol Vis Sci},
	volume = {54},
	number = {10},
	year = {2013},
	month = {2013 Oct 09},
	pages = {6612-9},
	abstract = {PURPOSE: We have previously shown that TGF-β3 (T3) stimulates extracellular matrix (ECM) assembly while maintaining antifibrotic characteristics in a model using human corneal fibroblasts (HCFs). This model, however, requires non-physiological levels of serum. In the current study, we tested whether T3 could stimulate human corneal keratocytes (HCKs) in vitro to assemble a functional ECM, while maintaining their characteristics. METHODS: Human corneal keratocytes and HCFs were isolated and cultured using 1\% or 10\% serum, respectively {\textpm}T3. The constructs were processed for indirect immunofluorescence (IF), transmission electron microscopy (TEM), and qRT-PCR, analyzing for keratocyte marker, keratocan, and ECM components, collagen (col) types I, III, and V. RESULTS: Quantitative reverse transcriptase PCR data showed that keratocan, col I, and V were all upregulated in HCKs compared with HCFs, whereas col III was expressed at low levels in HCKs. Transforming growth factor beta 3 stimulation further enhanced the level of change. Without T3, HCK constructs were very thin, approximately 5 μm; however, as with HCFs, upon stimulation with T3, HCK constructs increased in thickness by approximately 5-fold. Cell counts and ECM production revealed that HCKs assembled more ECM per unit area compared with HCFs, and IF revealed downregulation of fibrotic markers, col III, and thrombospondin-1, with T3 stimulation. Transmission electron microscopy data revealed aligned ECM with long fibrils for all conditions except HCK Controls. Human corneal keratocytes+T3 also showed denser collagen fibrils with more consistent fibril diameter. CONCLUSIONS: Overall, the data suggests that it is possible to stimulate matrix secretion and assembly by HCKs in vitro by using a single growth factor, T3.},
	keywords = {Biomarkers, Cells, Cultured, Corneal Keratocytes, Extracellular Matrix, Fibroblasts, Humans, Microscopy, Electron, Transmission, Reverse Transcriptase Polymerase Chain Reaction, Transforming Growth Factor beta3},
	issn = {1552-5783},
	doi = {10.1167/iovs.13-12861},
	author = {Karamichos, Dimitrios and Rich, Celeste B and Zareian, Ramin and Hutcheon, Audrey E K and Ruberti, Jeffrey W and Trinkaus-Randall, Vickery and Zieske, James D}
}
