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

被引:14
作者
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 ]
机构
[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
基金
巴西圣保罗研究基金会;
关键词
cellular reprogramming; induced pluripotent stem cells (iPSC); neural progenitor cells; swine; HUMAN FIBROBLASTS; DIRECTED DIFFERENTIATION; NEURONS; DISEASE; NEUROGENESIS; INDUCTION; MUTATION; PIG;
D O I
10.1002/term.3143
中图分类号
Q813 [细胞工程];
学科分类号
摘要
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.
引用
收藏
页码:1880 / 1891
页数:12
相关论文
共 46 条
[1]   Hepatic differentiation of porcine induced pluripotent stem cells in vitro [J].
Aravalli, Rajagopal N. ;
Cressman, Erik N. K. ;
Steer, Clifford J. .
VETERINARY JOURNAL, 2012, 194 (03) :369-374
[2]   Generation of induced pluripotent stem cells from large domestic animals [J].
Bressan, Fabiana Fernandes ;
Bassanezze, Vinicius ;
de Figueiredo Pessoa, Lais Vicari ;
Sacramento, Chester Bittencourt ;
Malta, Tathiane Maistro ;
Kashima, Simone ;
Fantinato Neto, Paulo ;
Strefezzi, Ricardo De Francisco ;
Godoy Pieri, Naira Caroline ;
Krieger, Jose Eduardo ;
Covas, Dimas Tadeu ;
Meirelles, Flavio Vieira .
STEM CELL RESEARCH & THERAPY, 2020, 11 (01) :247
[3]   Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling [J].
Chambers, Stuart M. ;
Fasano, Christopher A. ;
Papapetrou, Eirini P. ;
Tomishima, Mark ;
Sadelain, Michel ;
Studer, Lorenz .
NATURE BIOTECHNOLOGY, 2009, 27 (03) :275-280
[4]   Epithelial to mesenchymal transition-related proteins ZEB1, β-catenin, and β-tubulin-III in idiopathic pulmonary fibrosis [J].
Chilosi, Marco ;
Calio, Anna ;
Rossi, Andrea ;
Gilioli, Eliana ;
Pedica, Federica ;
Montagna, Licia ;
Pedron, Serena ;
Confalonieri, Marco ;
Doglioni, Claudio ;
Ziesche, Rolf ;
Grubinger, Markus ;
Mikulits, Wolfgang ;
Poletti, Venerino .
MODERN PATHOLOGY, 2017, 30 (01) :26-38
[5]   Renewed focus on the developing human neocortex [J].
Clowry, Gavin ;
Molnar, Zoltan ;
Rakic, Pasko .
JOURNAL OF ANATOMY, 2010, 217 (04) :276-288
[6]   Niche-independent symmetrical self-renewal of a mammalian tissue stem cell [J].
Conti, L ;
Pollard, SM ;
Gorba, T ;
Reitano, E ;
Toselli, M ;
Biella, G ;
Sun, YR ;
Sanzone, S ;
Ying, QL ;
Cattaneo, E ;
Smith, A .
PLOS BIOLOGY, 2005, 3 (09) :1594-1606
[7]   Generation and miRNA Characterization of Equine Induced Pluripotent Stem Cells Derived from Fetal and Adult Multipotent Tissues [J].
de Figueiredo Pessoa, Lais Vicari ;
Lisboa Pires, Pedro Ratto ;
del Collado, Maite ;
Godoy Pieri, Naira Caroline ;
Recchia, Kaiana ;
Souza, Aline Fernanda ;
Perecin, Felipe ;
da Silveira, Juliano Coelho ;
Cesar de Andrade, Andre Furugen ;
Ambrosio, Carlos Eduardo ;
Bressan, Fabiana Fernandes ;
Meirelles, Flavio Vieira .
STEM CELLS INTERNATIONAL, 2019, 2019
[8]   Dynamics of male canine germ cell development [J].
de Souza, Aline F. ;
Godoy Pieri, Naira C. ;
Roballo, Kelly C. S. ;
Bressan, Fabiana F. ;
Casals, Juliana B. ;
Ambrosio, Carlos E. ;
Perecin, Felipe ;
Martins, Daniele S. .
PLOS ONE, 2018, 13 (02)
[9]   Differentiation of Porcine Inner Cell Mass Cells Into Proliferating Neural Cells [J].
du Puy, Leonie ;
Lopes, Susana M. Chuva de Sousa ;
Haagsman, Henk P. ;
Roelen, Bernard A. J. .
STEM CELLS AND DEVELOPMENT, 2010, 19 (01) :61-70
[10]   Pluripotent Stem Cells from Domesticated Mammals [J].
Ezashi, Toshihiko ;
Yuan, Ye ;
Roberts, R. Michael .
ANNUAL REVIEW OF ANIMAL BIOSCIENCES, VOL 4, 2016, 4 :223-+