cultivation and characterization of cornea limbal epithelial stem cells on lens capsule in animal material-free medium培养和鉴定的角膜缘的晶状体囊上皮干细胞在动物材料免费媒介.pdf

Cultivation and Characterization of Cornea Limbal Epithelial Stem Cells on Lens Capsule in Animal Material- Free Medium ´ 1,2. ´ ´ 3. ´ ´ 2 4 4 Reka Albert , Zoltan Vereb , Krisztian Csomos , Morten C. Moe , Erik O. Johnsen , Ole 5 4 ´ ¨ 3 ´ ´ ´ ¨ 2 ´ 1 Kristoffer Olstad , Bjørn Nicolaissen , Eva Rajnavolgyi , Laszlo Fesus , Andras Berta , Goran Petrovski1,2* 1 Department of Ophthalmology, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary, 2 Stem Cells and Eye Research Laboratory, Department of Biochemistry and Molecular Biology, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary, 3 Department of Immunology, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary, 4 Centre of Eye Research, Department of Ophthalmology, Oslo University Hospital and University of Oslo, Oslo, Norway, 5 Department of Medical Biochemistry, Oslo University Hospital and University of Oslo, Oslo, Norway Abstract A simple, reproducible, animal-material free method for cultivating and characterizing cornea limbal epithelial stem cells (LESCs) on human lens capsule (LC) was developed for future clinical transplantation. The limbal tissue explants (2 62 60.25 mm) were harvested from 77 cadavers and expanded ex vivo on either cell culture plates or LC in medium containing human serum as the only growth supplement. Cell outgrowth at the edge of the explants was observed within 24 hours of cultivation and achieved viable outgrowth (.97% viability as measured by MTT assay and flow cytometry) within two weeks. The outgrowing cells were examined by gen