Culturing Human Bone Marrow-Derived Mesenchymal Stem Cells on Nanoskin Scaffolds for Ocular Surface Reconstruction.

• Grant number: 15/10727-0
• Support Opportunities: Regular Research Grants
• Start date: March 01, 2016
• End date: August 31, 2018
• Field of knowledge: Health Sciences - Medicine - Surgery
• Principal Investigator: Silvana Artioli Schellini
• Grantee: Silvana Artioli Schellini
• Host Institution: Faculdade de Medicina (FMB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
• Associated researchers: Alexandre Todorovic Fabro ; Carlos Roberto Padovani ; Claudia Aparecida Rainho ; Gabriel Molina de Olyveira ; Magda Massae Hata Viveiros ; Márcia Guimarães da Silva ; Márcio Eduardo de Melo Viveiros ; Nick Di Girolamo ; Pierre Basmaji ; Ramon Kaneno ; Valdecir Farias Ximenes

Abstract

Purpose: To establish the culture of mesenchymal stem cells derived from human bone marrow and access the properties of Nanoskin as scaffold for ex vivo cell transplantation in rabbit corneas.Methods: The bacterial cellulose three-dimensional fibrils scaffold in nanoscale (Nanoskin), will be studied as its morphology in scanning electron microscopy, as the optical transparency in a spectrophotometer and as the immunogenicity by ELISA study of the inflammatory cytokines produced by lymphocytes seeded over the nanoskin in addiction to lymphoproliferation assay with succinimidyl ester diacetate carboxyfluorescein (CFSE). The mesenchymal stem cells (MSCs) obtained from human bone marrow will be cultured until the second passage when its characterization will be accessed by flow cytometry. MSCs will be induced to differentiate into cornea epithelial cells through the contact with conditioned medium from corneal limbal cell cultures, with subsequent study using flow cytometry to confirm their transdifferentiation. MSCs undifferentiated and differentiated into keratinocytes will be seeded on Nanoskin and will be studied to access the cell adhesion in scanning electron microscopy and for cell viability by MTT test. The scaffold with the MSCs will be transferred onto wounded rabbit corneas in organ culture bath in order to study the cell integration, that will be accessed by immuno staining using antihuman nuclear antibody marker to identify human cells reepithelializating the lesions. (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)

MAGDA MASSAE HATA VIVEIROS; LUÍS HENRIQUE ZUCOLOTO; ÁLVIO ISSAO SHIGUEMATSU; CLÁUDIA APARECIDA RAINHO; SILVANA ARTIOLI SCHELLINI. Comparação das técnicas de cultivo de células epiteliais da córnea para a coleta de meio condicionado. Arquivos Brasileiros de Oftalmologia, v. 87, n. 2, 2024-04-22. (15/10727-0)

HATA VIVEIROS, MAGDA MASSAE; RAINHO, CLAUDIA APARECIDA; ZAMAME RAMIREZ, JOFER ANDREE; KANENO, RAMON; SILVA, MARCIA GUIMARAES; XIMENES, VALDECIR FARIAS; DE OLYVEIRA, GABRIEL MOLINA; BASMAJI, PIERRE; DI GIROLAMO, NICK; SCHELLINI, SILVANA ARTIOLI. Physical, functional and biochemical features of Nanoskin (R) bacterial cellulose scaffold as a potential carrier for cell transference. Materials Letters, v. 308, n. A, FEB 1 2022. (15/10727-0)