Generation of neural progenitor cells from porcine-induced pluripotent stem cells

Author(s):Show less -

Machado, Lucas Simoes ^{[1, 2]} ; Pieri, Naira Caroline Godoy ^[3] ; Botigelli, Ramon Cesar ^[4] ; de Castro, Raquel Vasconcelos Guimaraes ^[5] ; de Souza, Aline Fernanda ^[1] ; Bridi, Alessandra ^[1] ; Lima, Marina Amaro ^{[1, 2]} ; Fantinato Neto, Paulo ^[1] ; Pessoa, Lais Vicari de Figueiredo ^[1] ; Martins, Simone Maria Massami Kitamura ^[6] ; De Andrade, Andre Furugen Cesar ^[3] ; Freude, Kristine Karla ^[7] ; Bressan, Fabiana Fernandes ^{[1, 2]}

Affiliation

^[1] Univ Sao Paulo, Fac Anim Sci & Food Engn, Dept Vet Med, Pirassununga - Brazil

^[2] Univ Sao Paulo, Fac Vet Med & Anim Sci, Dept Surg, Postgrad Program Anat Domest & Wild Anim, Sao Paulo - Brazil

^[3] Univ Sao Paulo, Fac Vet Med & Anim Sci, Dept Anim Reprod, Sao Paulo - Brazil

^[4] Sao Paulo State Univ, Inst Biosci, Dept Pharmacol, Botucatu, SP - Brazil

^[5] Sao Paulo State Univ, Fac Agr & Vet Sci, Dept Prevent Vet Med & Anim Reprod, Jaboticabal - Brazil

^[6] Univ Sao Paulo, Fac Anim Sci & Food Engn, Dept Anim Sci, Pirassununga - Brazil

^[7] Univ Copenhagen, Dept Vet & Anim Sci, Copenhagen - Denmark

• Document type: Journal article
• Source: JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE; v. 14, n. 12 NOV 2020.
• Web of Science Citations: 0

Abstract

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, beta-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. (AU)

• FAPESP's process: 17/13973-8 - Generation and functional characterization of neuronal cells derived from iPSCs from patients with neurodegenerative diseases
• Grantee: Lucas Simões Machado
• Support Opportunities: Scholarships abroad - Research Internship - Master's degree
• FAPESP's process: 15/26818-5 - Investigation of cellular and molecular mechanisms on in vitro induced toti- and pluripotency acquisition - a translational approach
• Grantee: Fabiana Fernandes Bressan
• Support Opportunities: Research Grants - Young Investigators Grants
• FAPESP's process: 13/08135-2 - CTC - Center for Cell-Based Therapy
• Grantee: Dimas Tadeu Covas
• Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC
• FAPESP's process: 17/02159-8 - Induction of cell pluripotency and in vitro differentiation in the porcine model as a translational model
• Grantee: Lucas Simões Machado
• Support Opportunities: Scholarships in Brazil - Master