Cornea - ARVO

ARVO 2013 Annual Meeting Abstracts by Scientific Section/Group - CorneaConclusions: These findings demonstrated that periostin is a novelprotein that is mainly expressed by basal layer of human limbalepithelial cells and co-localized with p63. Periostin expression isassociated with higher proliferative capacity and less differentiationconditions in HCECs. Periostin may have a potential role inmaintaining the properties of human corneal epithelial progenitorcellsCommercial Relationships: Yangluowa Qu, None; Cheng Li,None; Wei Li, None; Zuguo Liu, Bausch&Lomb (R), Allergern (R),Alcon (R), Santen (R)Support: supported by Chinese National key sencientific researchproject 2013CB967003Program Number: 981 Poster Board Number: B0286Presentation Time: 1:00 PM - 2:45 PMDifferentiation Potential of Limbal Fibroblasts and BoneMarrow Mesenchymal Stem Cells to Corneal Epithelial CellsKishore Reddy Katikireddy, Reza Dana, Ula V. Jurkunas. SchepensEye Research Institute and Massachusetts Eye and Ear, Departmentof Ophthalmology, Harvard Medical School., Boston, MA.Purpose: It has been shown that human limbal fibroblasts (LFs) andbone marrow mesenchymal stem cells (BM MSCs) are multipotent.Specifically, a side population of stage-specific embryonic antigen-4(SSEA4)-positive LFs differentiates into a variety of cell types. Thepresent study compared the stem cell characteristics of SSEA4+ andSSEA4- LFs to those of BM MSCs, and determined their potential todifferentiate into the corneal epithelial phenotype.Methods: Human cadaveric limbal tissue (n=6) was treated withdispase to remove the epithelium and endothelium. Single cellsuspensions were obtained by digesting the stroma with 0.025%trypsin. Stem cell enrichment was performed by exposure of BMMSCs and LF cells in KnockOut ESC/iPSC medium for 12-15 days.LF and BM MSCs were sorted for SSEA4+ and SSEA4- cells byMagnetic-Activated Cell Sorting. Cell doubling time (CDT), stemcell (SC) marker analysis, and colony forming efficacy (CFE) wereperformed on sorted LF and BM MSCs. Epithelial phenotype wasachieved using induction and differentiation media. Differentiatedcells were characterized for corneal cytokeratins (CKs).Results: After separation, enriched LFs and BM MSCs, 97.4 ± 0.6%and 93.5 ± 0.7% SSEA4+ subgroups were achieved, respectively.The CDT of SSEA4+ LFs was 102 ± 1 hr and SSEA4- LFs was 58.2± 1.5 hrs (p=0.002). CDT of SSEA4+ BM MSCs was 105 ± 1 hr. andSSEA4- BM MSC was 56.3 ± 2 hrs (p=0.002). LF and BM MSCsubgroups were negative for pan-cytokeratin. After enrichment,SSEA4+ cells showed the ability to form cell aggregates, whileSSEA4- cells were mostly adherent to the culture plates. Thetranscript levels of SC markers OCT4, SOX2, Nanog and Rexo1were higher in SSEA4+ than SSEA4- groups of LF and BM MSCs(p