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Transplantation of amniotic membrane with limbal epithelial cells cultivated in sandwich configuration: clinical study and viability in rabbit corneas

Grant number: 14/18007-4
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): November 01, 2014
Effective date (End): September 30, 2017
Field of knowledge:Agronomical Sciences - Veterinary Medicine
Cooperation agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
Principal Investigator:José Luiz Laus
Grantee:Karina Kamachi Kobashigawa
Home Institution: Faculdade de Ciências Agrárias e Veterinárias (FCAV). Universidade Estadual Paulista (UNESP). Campus de Jaboticabal. Jaboticabal , SP, Brazil


This project was designed aiming the study of a technique for epithelial limbal stem cells culture and its therapeutic use. Limbal explants will be collected from 20 New Zealand rabbits and processed as cell suspensions, they will be used in two groups: Group I - epithelial cells expanded on a single layer of amniotic membrane. Group II - epithelial cells cultured in a tridimensional system composed by two layers of amniotic membrane (sandwich configuration). These samples will be transplanted in rabbits eyes with limbal stem cell deficiency and evaluated 1,3, 7, 14, 21, 28, 35, 42 e 45 after surgery. Blepharospasm, ocular secretion, conjunctival hyperemia, quemosis, corneal vascularization and transparency will be characterized by its intensity. Immunochemistry will be employed to study p63 transcription factor, Ki67 cellular proliferation factor and cytokeratins 3, 12 and 19, in days 7 and 45 after transplantation. Parameters with normal distribution will be tested with t Student test and non normal with Mann-Whitney test. Statistical difference will be considered when pÂ0.05. (AU)

Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
KOBASHIGAWA, KARINA KAMACHI; ALDROVANI, MARCELA; BARROS SOBRINHO, ALEXANDRE A. F.; SANTO SILVA, PALOMA DO ESPIRITO; MARCUSSO, PAULO F.; MARINHO-NETO, FAUSTO A.; MARTINES PADUA, IVAN R.; LAUS, JOSE LUIZ. Clinical features and corneal optical anisotropies in a rabbit model of limbal stem cell deficiency. Arquivos Brasileiros de Oftalmologia, v. 81, n. 5, p. 384-392, SEP-OCT 2018. Web of Science Citations: 0.
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
KOBASHIGAWA, Karina Kamachi. Transplante de membrana amniótica com células epiteliais limbais cultivadas em sanduíche : estudos clínicos e de viabilidade em córneas de coelhos. 2018. Doctoral Thesis - Universidade Estadual Paulista "Júlio de Mesquita Filho" Faculdade de Ciências Agrárias e Veterinárias..

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