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Ilham Putra, Gaurav Agnihotri, Samaneh Ghassemi, Judy Hamouie, Asha Tadepalli, Xiang Shen, Peiman Hematti, Medi Eslani, Ali R Djalilian; Human Corneal Limbal Stromal Cells provide a source of Mesenchymal Stem Cells. Invest. Ophthalmol. Vis. Sci. 2016;57(12):892.
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© ARVO (1962-2015); The Authors (2016-present)
Mesenchymal stem cell (MSC)-based therapy is under investigation as a treatment strategy to promote tissue repair and to modulate the immune system in immune mediated disorders. MSCs have been isolated from many tissues most commonly from the bone marrow or fat. Given the cumbersome procedures for harvesting bone marrow (BM), and to a lesser extent fat, there is a need to look for other tissues for MSC isolation. The International Society for Cellular Therapy (ISCT) has established three criteria for characterizing human MSCs: adherence to plastic, differentiation potential, and the presence or absence of certain cell surface markers. We evaluated whether or not corneal limbal stromal cells (CLSC) met ISCT criteria by comparing them to BM-MSCs.
BM-MSCs were isolated from healthy human bone marrow donors. CLSC were isolated from human corneal limbus from research cadaver corneas (generously provided by Eversight). Both cell types were cultured in MEM- alpha + 10% fetal bovine serum. They were photographed for morphology at each passage. Cells from passage 4 were used for immunophenotyping using fluorescent-activated cell sorting (BD LSR Fortessa). Their differentiation into the three mesenchymal lineages (adipocytes, osteoblasts, and chondrocytes) was evaluated using staining with LipidTOX™ for fatty acids, Alizarin Red for calcium, and Alcian blue for glycosaminoglycans, respectively. All experiments were repeated with at least five different donors.
CLSCs and BM-MSCs were both able to adhere to the plastic. They had identical morphology. They were positive for the cell surface markers CD90, CD105, CD73 and negative for CD34, CD45, HLA-DR, CD19, and CD11b. After incubation in differentiation media, they were able to differentiate successfully into adipocytes, osteoblasts, and chondrocytes.
Based on ISCT criteria, CLSCs can be considered MSCs. They can be used as a convenient source of MSC for the cell-based therapy as they can be obtained from corneas from eye bank.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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