Isolation and characterization of mesenchymal stem cells derived from bovine Wharton's jelly and their potential for use in cloning by nuclear transfer.

SILVA, C. G. da; MARTINS, C. F.; CARDOSO, T. C.; CUNHA, E. R. da; CUMPA, H. C. B.; MCMANUS, C. M.; PIVATO, I.; BAO, S. N.

Please use this identifier to cite or link to this item: http://www.alice.cnptia.embrapa.br/alice/handle/doc/1051545

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DC Field	Value	Language
dc.contributor.author	SILVA, C. G. da	pt_BR
dc.contributor.author	MARTINS, C. F.	pt_BR
dc.contributor.author	CARDOSO, T. C.	pt_BR
dc.contributor.author	CUNHA, E. R. da	pt_BR
dc.contributor.author	CUMPA, H. C. B.	pt_BR
dc.contributor.author	MCMANUS, C. M.	pt_BR
dc.contributor.author	PIVATO, I.	pt_BR
dc.contributor.author	BAO, S. N.	pt_BR
dc.date.accessioned	2016-08-24T11:11:11Z	pt_BR
dc.date.available	2016-08-24T11:11:11Z	pt_BR
dc.date.created	2016-08-24	pt_BR
dc.date.issued	2016	pt_BR
dc.identifier.citation	Ciência Rural, Santa Maria, v. 46, n. 10, p. 1830-1837, out. 2016.	pt_BR
dc.identifier.issn	ISSN 1678-4596	pt_BR
dc.identifier.uri	http://www.alice.cnptia.embrapa.br/alice/handle/doc/1051545	pt_BR
dc.description	RESUMO: A geleia de Wharton é uma fonte de células tronco mesenquimais (CTMs) que ainda não havia sido testada para a produção de embriões bovinos por transferência nuclear (TN). O objetivo deste estudo foi isolar, caracterizar e testar as CTMs derivadas da geleia de Wharton para produção de embriões e gestações por transferência nuclear em bovinos. O cordão umbilical foi coletado durante o nascimento e as células derivadas da geleia de Wharton (CGWs) foram isoladas por explante e cultivadas em Dulbecco?s Modified Eagle Medium. Fibroblastos (FB) da pele foram isolados após 6 meses de vida. As análises morfológicas foram realizadas pelas microscopias de campo claro e eletrônica de varredura durante o cultivo celular. Caracterização fenotípica e genotípica por citometria de fluxo, imunocitoquímica, RT-PCR e indução da diferenciação em linhagens celulares foi realizada com as CGWs. No procedimento de TN, ovócitos no estágio de metáfase II foram enucleados usando micromanipuladores, fusionados com CGWs ou FB e então ativados artificialmente. Micrografias de microscopia de varredura revelaram que CGWs tiveram forma variada sob cultivo. Os marcadores mesenquimais de CTMs (CD29+, CD73+, CD90+ and CD105+) foram expressos em cultura de CGWs bovina, como evidenciado por citometria de fluxo, imunocitoquímica e RT-PCR. Quando induzidas, estas células diferenciaram-se em osteócitos, condrócitos e adipócitos. Após classificação, as CGWs foram utilizadas na TN. A taxa de formação de blastocistos por TN com CGWs no sétimo dia de cultivo foi de 25,80±0,03%, similar a produção de blastócitos por TN com fibroblastos de pele (19,00±0,07). Gestações foram obtidas e mostraram que CGWs constituem um novo tipo celular para ser usado na clonagem animal. ABSTRACT: Wharton?s jelly is a source of mesenchymal stem cells (MSCs) that had not yet been tested for bovine embryo production by nuclear transfer (NT). Thus, the objective of this study was to isolate, characterize and test MSCs derived from Wharton?s jelly for embryo and pregnancy production by NT in cattle. The umbilical cord was collected during calving and cells derived from Wharton?s jelly (WJCs) were isolated by explant and cultured in Dulbecco?s Modified Eagle Medium. Skin Fibroblasts (FB) were isolated after 6 months of life. Morphological analysis was performed by bright field and scanning electron microscopy (SEM) during cell culture. Phenotypic and genotypic characterization by flow cytometry, immunocytochemistry, RT-PCR and differentiation induction in cell lineages were performed for WJC. In the NT procedure, oocytes at the arrested metaphase II stage were enucleated using micromanipulators, fused with WJCs or FB and later activated artificially. SEM micrographs revealed that WJCs have variable shape under culture. Mesenchymal markers of MSCs (CD29+, CD73+, CD90+ and CD105+) were expressed in bovine-derived WJC cultures, as evidenced by flow cytometry, immunocytochemistry and RT-PCR. When induced, these cells differentiated into osteocytes, chondrocytes and adipocytes. After classification, the WJCs were used in NT. Blastocyst formation rate by NT with WJCs at day 7 was 25.80±0.03%, similar to blatocyst rate with NT using skin fibroblasts (19.00±0.07%). Pregnancies were obtained and showed that WJCs constitute a new cell type for use in animal cloning.	pt_BR
dc.language.iso	eng	eng
dc.rights	openAccess	eng
dc.subject	Célula tronco mesenquimal	pt_BR
dc.subject	Cordão umbilical	pt_BR
dc.subject	Geleia de Wharton	pt_BR
dc.subject	Reprodução assistida	pt_BR
dc.title	Isolation and characterization of mesenchymal stem cells derived from bovine Wharton's jelly and their potential for use in cloning by nuclear transfer.	pt_BR
dc.type	Artigo de periódico	pt_BR
dc.date.updated	2016-09-08T11:11:11Z	pt_BR
dc.subject.thesagro	Clonagem	pt_BR
dc.subject.thesagro	Reprodução Animal	pt_BR
riaa.ainfo.id	1051545	pt_BR
riaa.ainfo.lastupdate	2016-09-08	pt_BR
dc.identifier.doi	http://dx.doi.org/10.1590/0103-8478cr20151074	pt_BR
dc.contributor.institution	CAROLINA GONZALES DA SILVA, UNB; CARLOS FREDERICO MARTINS, CPAC; TEREZA CRISTINA CARDOSO, UNESP; ELISA RIBEIRO DA CUNHA, UNB; HEIDI CHRISTINA BESSLER CUMPA, CPAC; CONCEPTA MARGARET MCMANUS, UNB; IVO PIVATO, UNB; SÔNIA NAIR BÁO, UNB.	pt_BR
Appears in Collections:	Artigo em periódico indexado (CPAC)

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