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Induction of Corneal Epithelial Differentiation of Induced Pluripotent and Orbital Fat-Derived Stem Cells Seeded on Decellularized Human Corneas.
da Mata Martins, Thaís Maria; de Carvalho, Juliana Lott; da Silva Cunha, Pricila; Gomes, Dawidson Assis; de Goes, Alfredo Miranda.
Afiliação
  • da Mata Martins TM; Department of Morphology, Institute of Biological Sciences, Federal University of Minas Gerais, Avenida Presidente Antônio Carlos, 6627, Belo Horizonte, Minas Gerais, 31270-901, Brazil. thaismmmartins@gmail.com.
  • de Carvalho JL; Department of Genomic Sciences and Biotechnology, Catholic University of Brasilia, QS 07 - Lote 01, EPCT - Taguatinga, Brasília, Distrito Federal, 71966-700, Brazil.
  • da Silva Cunha P; Faculty of Medicine, University of Brasilia, Campus Universitário Darcy Ribeiro, Brasília, Distrito Federal, 70910-900, Brazil.
  • Gomes DA; Department of Biochemistry and Immunology, Institute of Biological Sciences, Federal University of Minas Gerais, Avenida Presidente Antônio Carlos, 6627, Belo Horizonte, Minas Gerais, 31270-901, Brazil.
  • de Goes AM; Department of Biology, Minas Gerais State University, Avenida Olegário Maciel, 1427, Ubá, Minas Gerais, 36502-002, Brazil.
Stem Cell Rev Rep ; 18(7): 2522-2534, 2022 10.
Article em En | MEDLINE | ID: mdl-35247143
Up to 40% of donor corneas are deemed unsuitable for transplantation, aggravating the shortage of graft tissue. In most cases, the corneal extracellular matrix is intact. Therefore, their decellularization followed by repopulation with autologous cells may constitute an efficient alternative to reduce the amount of discarded tissue and the risk of immune rejection after transplantation. Although induced pluripotent (hiPSCs) and orbital fat-derived stem cells (OFSCs) hold great promise for corneal epithelial (CE) reconstruction, no study to date has evaluated the capacity of decellularized corneas (DCs) to support the attachment and differentiation of these cells into CE-like cells. Here, we recellularize DCs with hiPSCs and OFSCs and evaluate their differentiation potential into CE-like cells using animal serum-free culture conditions. Cell viability and adhesion on DCs were assessed by calcein-AM staining and scanning electron microscopy. Cell differentiation was evaluated by RT-qPCR and immunofluorescence analyses. DCs successfully supported the adhesion and survival of hiPSCs and OFSCs. The OFSCs cultured under differentiation conditions could not express the CE markers, TP63, KRT3, PAX6, and KRT12, while the hiPSCs gave rise to cells expressing high levels of these markers. RT-qPCR data suggested that the DCs provided an inductive environment for CE differentiation of hiPSCs, supporting the expression of PAX6 and KRT12 without the need for any soluble induction factors. Our results open the avenue for future studies regarding the in vivo effects of DCs as carriers for autologous cell transplantation for ocular surface reconstruction.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Tecido Adiposo / Células-Tronco Pluripotentes Induzidas Limite: Animals / Humans Idioma: En Ano de publicação: 2022 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Tecido Adiposo / Células-Tronco Pluripotentes Induzidas Limite: Animals / Humans Idioma: En Ano de publicação: 2022 Tipo de documento: Article