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Generation of neural progenitor cells from porcine‐induced pluripotent stem cells
Author(s) -
Machado Lucas Simões,
Pieri Naira Caroline Godoy,
Botigelli Ramon Cesar,
Castro Raquel Vasconcelos Guimarães,
Souza Aline Fernanda,
Bridi Alessandra,
Lima Marina Amaro,
Fantinato Neto Paulo,
Pessôa Laís Vicari de Figueiredo,
Martins Simone Maria Massami Kitamura,
De Andrade André Furugen Cesar,
Freude Kristine Karla,
Bressan Fabiana Fernandes
Publication year - 2020
Publication title -
journal of tissue engineering and regenerative medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.835
H-Index - 72
eISSN - 1932-7005
pISSN - 1932-6254
DOI - 10.1002/term.3143
Subject(s) - sox2 , embryoid body , nestin , neural stem cell , embryonic stem cell , induced pluripotent stem cell , homeobox protein nanog , neurosphere , microbiology and biotechnology , klf4 , biology , progenitor cell , stem cell , vimentin , adult stem cell , immunology , immunohistochemistry , biochemistry , gene
Summary In this study, porcine embryonic fibroblasts (pEFs) were reprogrammed into porcine‐induced pluripotent stem cells (piPSCs) using either human or mouse specific sequences for the OCT4 , SOX2 , c‐Myc , and KLF4 transcription factors. In total, three pEFs lines were reprogrammed, cultured for at least 15 passages, and characterized regarding their pluripotency status (alkaline phosphatase expression, embryoid body formation, expression of exogenous and endogenous genes, and immunofluorescence). Two piPSC lines were further differentiated, using chemical inhibitors, into putative neural progenitor‐like (NPC‐like) cells with subsequent analyses of their morphology and expression of neural markers such as NESTIN and GFAP as well as immunofluorescent labeling of NESTIN, β‐TUBULIN III, and VIMENTIN. NPC‐like cells were positive for all the neural markers tested. These results evidence of the generation of porcine NPC‐like cells after in vitro induction with chemical inhibitors, representing an adequate model for future regenerative and translational medicine research.