Cornea - ARVO

ARVO 2013 Annual Meeting Abstracts by Scientific Section/Group - Corneamarkedly decline based on DiO labeling. This suggests that techniquemodifications may be required to optimize intra-stromal stem cellviability. Antibody staining for markers of human stem celltransplantation and keratocyte differentiation may also aid ourunderstanding of corneal stem cell fate and verify loss of cells orlabeling.Commercial Relationships: Behdad Kavianpour, None; GeraintJ. Parfitt, None; Hongshan Liu, None; Winston W. Kao, None;Donald J. Brown, None; Yilu Xie, None; Mikhail Geyfman, None;James V. Jester, None; Jennifer Simpson, NoneSupport: NIH Grant EY022365 Mesenchymal Stem Cell TherapyFor Corneal Cystinosis; Research to Prevent Blindness, Inc;Discovery Eye Foundation; The Skirball Program in MolecularOphthalmology; The Cystinosis Research FoundationProgram Number: 1013 Poster Board Number: B0318Presentation Time: 1:00 PM - 2:45 PMInfluence of Secreted Ly6/uPAR-Related Protein-1 (Slurp1) onCorneal Stromal Fibroblast Proliferation, Interaction with theExtracellular Matrix, and MigrationShivalingappa K. Swamynathan 1, 2 , Sudha Swamynathan 1 .1 Ophthalmology, Univ Pittsburgh Sch of Med, Pittsburgh, PA; 2 CellBiology and Physiology, Univ Pittsburgh Sch of Med, Pittsburgh,PA.Purpose: Previously, we demonstrated that the secreted Ly6/uPARrelatedprotein-1 (Slurp1) is abundantly expressed in the cornea andis downregulated in diverse pro-inflammatory conditions. Here, weexamine the effects of Slurp1 on corneal stromal fibroblast cellproliferation, interaction with the extracellular matrix (ECM), andmigration, to understand the cellular basis of Slurp1 functions in thecornea.Methods: The effect of Slurp1 on corneal fibroblast behavior wasassessed in vitro by treating human telomerase reverse transcriptase(hTERT)-immortalized mouse corneal stromal cell line MK/T1 withhistidine-tagged mouse Slurp1 (His-Slurp1) produced in E. coli andpartially purified by Ni ion resin column chromatography. Effect ofHis-Slurp1 on MK/T1 cell (i) density was assessed by crystal violetstaining followed by measurement of absorbance at 590 nm, (ii)interaction with the ECM was evaluated on cell culture plates coatedwith different ECM components, and (iii) migration was assessed byin vitro gap filling assays.Results: Compared with the control, His-Slurp1-treated mousecorneal stromal fibroblast MK/T1 cells (i) density increased at aslower pace suggesting that Slurp1 inhibits cell proliferation, (ii)adhered with lower affinity to collagen-I-, collagen-IV-, vitronectinorfibronectin-coated culture plates suggesting that Slurp1 affectscell-matrix interaction, and (iii) migrated at a slower pace in gapfilling assays suggesting that Slurp1 inhibits cell migration.Conclusions: Our results demonstrate that Slurp1 inhibits cornealstromal fibroblast MK/T1 cell proliferation, cell-matrix adhesion andmigration, revealing the cellular basis for corneal functions of Slurp1.These results are consistent with the decreased expression of Slurp1in corneas exposed to pro-inflammatory conditions where the stromalfibroblasts proliferate at a higher rate, and migrate rapidly.Commercial Relationships: Shivalingappa K. Swamynathan,None; Sudha Swamynathan, NoneSupport: Department of Ophthalmology, University of PittsburghStart-up funds, Eye and Ear Foundation of Pittsburgh, Research toPrevent Blindness and NIH core grant P30 EY08098Program Number: 1014 Poster Board Number: B0319Presentation Time: 1:00 PM - 2:45 PMStromal Cells derived from Amniotic Membrane are capable toreestablish corneal opacityYonathan Garfias 1, 2 , Alejandro Navas 1 , Jessica Nieves-Hernández 1 ,Gibran A. Estua 1 , Rodrigo Bolaños-Jiménez 1 . 1 Research Unit,Institute of Ophthalmology, Mexico City, Mexico; 2 Biochemistry,Faculty of Medicine, Universidad Nacional Autónoma de México,Mexico City, Mexico.Purpose: Stromal mesenchymal stem cells are non-hematopoieticderived cells found in the bone marrow stroma such as in manystromal tissues. The amniotic membrane is an elastin and avascularfetal membrane that is in contact to the fetus. A mature amnioticmembrane possesses 20-50 x 10 mesenchymal cells. .By the other hand, there are many corneal disorders that directlyaffect the corneal limbus, driving inflammation, conjunctivalizationor neovascularization of the corneal tissue. The pronostic depends onthe injured area of the limbus where the corneal stem cells arelocalized. Although, it has recently reported that the cells derivedfrom the amniotic membrane mesenchyma are source toadipogenesis, chondrogenesis, osteogenesis and myogenesis, itsfunction as a source for regeneration of the ocular surface has notbeen studied.The aim of the present study is to determine the utility of these cellsto restablish the ocular surface in a chemical burn murine model.Methods: Mesenchymal cells were obtained from a placenta usingdispase/collagenase method. The cells were cultured andcharacterized by flow cytometry. Cellular transdifferentiation assayswere performed using conditioned media. A murine chemical burnwas performed in order to determine the efficacy of these cells torestablish the corneal clarity. Corneal histology was performed toidentify the incorporation of human mesenchymal cells in the murinecornea.Results: The cells obtained from the amniotic membranemesenchyma were capable to attach to the plastic wells showing afibroblast-like morphology. These cells presented mesenchymal stemcell markers such as CD29, CD73, CD44 and CD105, meanwhile,they were negative to CD45 and HLA-DR. Interestingly, these cellswere capable to differentiate into neurons and chondrocytes. Whenthese cells were intracamerally injected in a mouse burn model, thecorneal opacity was significantly reduced in comparison to theuntreated cornea. When the histology of the cornea was performed, itwas evident that the human amniotic membrane cells wereincorporated to the mouse cornea, reestablishing the structure of thecorneal tissue.Conclusions: The use of cells derived from the mesenchyma of theamniotic membrane is an important cell source to be used in theregenerative ophthalmology.Commercial Relationships: Yonathan Garfias, Institute ofOphthalmology (P); Alejandro Navas, None; Jessica Nieves-Hernández, Institute of Ophthalmology (P); Gibran A. Estua,None; Rodrigo Bolaños-Jiménez, NoneSupport: CONACYT 160286Program Number: 1015 Poster Board Number: B0320Presentation Time: 1:00 PM - 2:45 PMCorneal endothelial cells derived from monkey iPS cells: a shortterm evaluationShin Hatou 1 , Satoru Yoshida 1 , Kazunari Higa 2 , Hideyuki Miyashita 1 ,Emi Inagaki 1 , Erika Kimura 3 , Ryuhei Hayashi 3 , Kazuo Tsubota 1 ,Kohji Nishida 3 , Shigeto Shimmura 1 . 1 Department of Ophthalmology,Keio Univ School of Medicine, Shinjuku-ku, Japan; 2 Department ofOphthalmology, Tokyo Dental College Ichikawa General Hospital,Ichikawa, Japan; 3 Department of Ophthalmology, Osaka University,Osaka, Japan.©2013, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permissionto reproduce any abstract, contact the ARVO Office at arvo@arvo.org.