Characterization and Immunomodulation of Canine Amniotic Membrane Stem Cells

Alessandra de Oliveira Pinheiro,1 Valéria M Lara,1 Aline F Souza,1 Juliana B Casals,2 Fabiana F Bressan,1 Paulo Fantinato Neto,1 Vanessa C Oliveira,1 Daniele S Martins,1 Carlos E Ambrosio1 1Department of Veterinary Medicine, Faculty of Animal Science and Food Engineering, University of S&...

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Autores principales:	de Oliveira Pinheiro A, Lara VM, Souza AF, Casals JB, Bressan FF, Fantinato Neto P, Oliveira VC, Martins DS, Ambrosio CE
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2020
Materias:	canine stem cells immunomodulation fetal annexes. Cytology QH573-671
Acceso en línea:	https://doaj.org/article/33ff5cbd2912475abaa07b3351b19ff4
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spelling	oai:doaj.org-article:33ff5cbd2912475abaa07b3351b19ff42021-12-02T09:54:47ZCharacterization and Immunomodulation of Canine Amniotic Membrane Stem Cells1178-6957https://doaj.org/article/33ff5cbd2912475abaa07b3351b19ff42020-05-01T00:00:00Zhttps://www.dovepress.com/characterization-and-immunomodulation-of-canine-amniotic-membrane-stem-peer-reviewed-article-SCCAAhttps://doaj.org/toc/1178-6957Alessandra de Oliveira Pinheiro,1 Valéria M Lara,1 Aline F Souza,1 Juliana B Casals,2 Fabiana F Bressan,1 Paulo Fantinato Neto,1 Vanessa C Oliveira,1 Daniele S Martins,1 Carlos E Ambrosio1 1Department of Veterinary Medicine, Faculty of Animal Science and Food Engineering, University of São Paulo, Pirassununga, São Paulo, Brazil; 2Private Veterinary Practice, Pirassununga, São Paulo, BrazilCorrespondence: Carlos E AmbrosioDepartment of Veterinary Medicine, Faculty of Animal Science and Food Engineering, University of São Paulo, FZEA- Av. Duque de Caxias Norte, 225, ZMV, Pirassununga 13635-900, São Paulo, BrazilTel +55 19 3565-4113 Email ceambrosio@usp.brPurpose: Amniotic membrane stem cells have a high capacity of proliferation, cell expansion, and plasticity, as well as immunomodulatory properties that contribute to maternal-fetal tolerance. Owing to the lack of research on human amniotic membrane at different gestational stages, the canine model is considered ideal because of its genetic and physiological similarities. We aimed to characterize the canine amniotic membrane (CAM) cell lineage in different gestational stages and evaluate the expression of immunomodulatory genes.Materials and Methods: Twenty CAMs from early (20– 30 days) (n=7), mid- (31– 45 days) (n=7), and late gestation (46– 63 days) (n=6) stages were studied. The cell features were assessed by cell viability tests, growth curve, colony-forming units, in vitro differentiation, cell labeling for different immunophenotypes, and pluripotent potential markers. The cells were subjected to RT-PCR and qPCR analysis to determine the expression of IDO, HGF, EGF, PGE2, and IL-10 genes.Results: CAM cells exhibited a fibroblastoid morphology and adherence to plastic with an average cell viability of 78.5%. The growth curve indicated a growth peak in the second passage and we obtained an average of 138.2 colonies. Osteogenic, chondrogenic, and adipogenic lineages were confirmed by in vitro differentiation assays. Cellular immunophenotyping experiments confirmed the presence of positive mesenchymal markers (CD90 and CD105) and the low or negative expression of hematopoietic markers (CD45 and CD34). Qualitative analysis of the immunomodulatory functions indicated the expression of the IDO, HGF, EGF5, and PGE2 genes. When stimulated by interferon-gamma, CAM cells exhibited higher IDO levels throughout gestation.Conclusion: The CAMs from different gestational stages presented features consistent with mesenchymal stem cell lineage; better results were observed during the late gestation stage. Therefore, the gestational stage is a key factor that may influence the functionality of therapies when using fetal membrane tissues from different periods of pregnancy.Keywords: canine stem cells, immunomodulation, fetal annexesde Oliveira Pinheiro ALara VMSouza AFCasals JBBressan FFFantinato Neto POliveira VCMartins DSAmbrosio CEDove Medical Pressarticlecanine stem cellsimmunomodulationfetal annexes.CytologyQH573-671ENStem Cells and Cloning: Advances and Applications, Vol Volume 13, Pp 43-55 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	canine stem cells immunomodulation fetal annexes. Cytology QH573-671
spellingShingle	canine stem cells immunomodulation fetal annexes. Cytology QH573-671 de Oliveira Pinheiro A Lara VM Souza AF Casals JB Bressan FF Fantinato Neto P Oliveira VC Martins DS Ambrosio CE Characterization and Immunomodulation of Canine Amniotic Membrane Stem Cells
description	Alessandra de Oliveira Pinheiro,1 Valéria M Lara,1 Aline F Souza,1 Juliana B Casals,2 Fabiana F Bressan,1 Paulo Fantinato Neto,1 Vanessa C Oliveira,1 Daniele S Martins,1 Carlos E Ambrosio1 1Department of Veterinary Medicine, Faculty of Animal Science and Food Engineering, University of São Paulo, Pirassununga, São Paulo, Brazil; 2Private Veterinary Practice, Pirassununga, São Paulo, BrazilCorrespondence: Carlos E AmbrosioDepartment of Veterinary Medicine, Faculty of Animal Science and Food Engineering, University of São Paulo, FZEA- Av. Duque de Caxias Norte, 225, ZMV, Pirassununga 13635-900, São Paulo, BrazilTel +55 19 3565-4113 Email ceambrosio@usp.brPurpose: Amniotic membrane stem cells have a high capacity of proliferation, cell expansion, and plasticity, as well as immunomodulatory properties that contribute to maternal-fetal tolerance. Owing to the lack of research on human amniotic membrane at different gestational stages, the canine model is considered ideal because of its genetic and physiological similarities. We aimed to characterize the canine amniotic membrane (CAM) cell lineage in different gestational stages and evaluate the expression of immunomodulatory genes.Materials and Methods: Twenty CAMs from early (20– 30 days) (n=7), mid- (31– 45 days) (n=7), and late gestation (46– 63 days) (n=6) stages were studied. The cell features were assessed by cell viability tests, growth curve, colony-forming units, in vitro differentiation, cell labeling for different immunophenotypes, and pluripotent potential markers. The cells were subjected to RT-PCR and qPCR analysis to determine the expression of IDO, HGF, EGF, PGE2, and IL-10 genes.Results: CAM cells exhibited a fibroblastoid morphology and adherence to plastic with an average cell viability of 78.5%. The growth curve indicated a growth peak in the second passage and we obtained an average of 138.2 colonies. Osteogenic, chondrogenic, and adipogenic lineages were confirmed by in vitro differentiation assays. Cellular immunophenotyping experiments confirmed the presence of positive mesenchymal markers (CD90 and CD105) and the low or negative expression of hematopoietic markers (CD45 and CD34). Qualitative analysis of the immunomodulatory functions indicated the expression of the IDO, HGF, EGF5, and PGE2 genes. When stimulated by interferon-gamma, CAM cells exhibited higher IDO levels throughout gestation.Conclusion: The CAMs from different gestational stages presented features consistent with mesenchymal stem cell lineage; better results were observed during the late gestation stage. Therefore, the gestational stage is a key factor that may influence the functionality of therapies when using fetal membrane tissues from different periods of pregnancy.Keywords: canine stem cells, immunomodulation, fetal annexes
format	article
author	de Oliveira Pinheiro A Lara VM Souza AF Casals JB Bressan FF Fantinato Neto P Oliveira VC Martins DS Ambrosio CE
author_facet	de Oliveira Pinheiro A Lara VM Souza AF Casals JB Bressan FF Fantinato Neto P Oliveira VC Martins DS Ambrosio CE
author_sort	de Oliveira Pinheiro A
title	Characterization and Immunomodulation of Canine Amniotic Membrane Stem Cells
title_short	Characterization and Immunomodulation of Canine Amniotic Membrane Stem Cells
title_full	Characterization and Immunomodulation of Canine Amniotic Membrane Stem Cells
title_fullStr	Characterization and Immunomodulation of Canine Amniotic Membrane Stem Cells
title_full_unstemmed	Characterization and Immunomodulation of Canine Amniotic Membrane Stem Cells
title_sort	characterization and immunomodulation of canine amniotic membrane stem cells
publisher	Dove Medical Press
publishDate	2020
url	https://doaj.org/article/33ff5cbd2912475abaa07b3351b19ff4
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Characterization and Immunomodulation of Canine Amniotic Membrane Stem Cells

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