title: Isolation and characterization of mesenchymal stem cells from the yolk sacs of bovine embryos.
abstract: The yolk sac (YS) represents a promising source of stem cells for research because of the hematopoietic and mesenchymal cell niches that are present in this structure during the development of the embryo. In this study, we report on the isolation and characterization of YS tissue and mesenchymal stem cells (MSCs) derived from bovine YSs. Our results show that the YS is macroscopically located in the exocoelomic cavity in the ventral portion of the embryo and consists of a transparent membrane formed by a central sac-like portion and two ventrally elongated projections. Immunohistochemistry analyses were positive for OCT4, CD90, CD105, and CD44 markers in the YS of both gestational age groups. The MSCs of bovine YS were isolated using enzymatic digestion and were grown in vitro for at least 11 passages to verify their capacity to proliferate. These cells were also subjected to immunophenotypic characterization that revealed the presence of CD90, CD105, and CD79 and the absence of CD45, CD44, and CD79, which are positive and negative markers of MSCs, respectively. To prove their multipotency, the cells were induced to differentiate into three cell types, chondrocytes, osteoblasts, and adipocytes, which were stained with tissue-specific dyes (chondrogenic: Alcian Blue, osteogenic: Alizarin Red, and adipogenic: Oil Red O) to confirm differentiation. Gene expression analyses showed no differences in the patterns of gene expression between the groups or passages tested, with the exception of the expression of SOX2, which was slightly different in the G1P3 group compared to the other groups. Our results suggest that YS tissue from bovines can be used as a source of MSCs, which makes YS tissue-derived cells an interesting option for cell therapy and regenerative medicine.
authors.affiliation: Department of Veterinary Medicine, Faculty of Animal Science and Food Engineering, University of São Paulo, Pirassununga, São Paulo, Brazil.
authors.forename: C A F
authors.initials: CA
authors.lastname: Mançanares
authors.affiliation: Department of Veterinary Medicine, Faculty of Animal Science and Food Engineering, University of São Paulo, Pirassununga, São Paulo, Brazil.
authors.forename: V C
authors.initials: VC
authors.lastname: Oliveira
authors.affiliation: Department of Veterinary Medicine, Faculty of Animal Science and Food Engineering, University of São Paulo, Pirassununga, São Paulo, Brazil.
authors.forename: L J
authors.initials: LJ
authors.lastname: Oliveira
authors.affiliation: Morphology Laboratory of Veterinary Medicine College, UNIFEOB, São João da Boa Vista, São Paulo, Brazil.
authors.forename: A F
authors.initials: AF
authors.lastname: Carvalho
authors.affiliation: Department of Surgery, Faculty of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, São Paulo, Brazil.
authors.forename: R V
authors.initials: RV
authors.lastname: Sampaio
authors.affiliation: Department of Surgery, Faculty of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, São Paulo, Brazil.
authors.forename: A C F
authors.initials: AC
authors.lastname: Mançanares
authors.affiliation: Department of Surgery, Faculty of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, São Paulo, Brazil.
authors.forename: A F
authors.initials: AF
authors.lastname: Souza
authors.affiliation: Department of Veterinary Medicine, Faculty of Animal Science and Food Engineering, University of São Paulo, Pirassununga, São Paulo, Brazil.
authors.forename: F
authors.initials: F
authors.lastname: Perecin
authors.affiliation: Department of Veterinary Medicine, Faculty of Animal Science and Food Engineering, University of São Paulo, Pirassununga, São Paulo, Brazil.
authors.forename: F V
authors.initials: FV
authors.lastname: Meirelles
authors.affiliation: Department of Surgery, Faculty of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, São Paulo, Brazil.
authors.forename: M A
authors.initials: MA
authors.lastname: Miglino
authors.affiliation: Department of Veterinary Medicine, Faculty of Animal Science and Food Engineering, University of São Paulo, Pirassununga, São Paulo, Brazil. Electronic address: ceambrosio@usp.br.
authors.forename: C E
authors.initials: CE
authors.lastname: Ambrósio
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002417:Cattle; D018929:Cell Culture Techniques / Q000662:veterinary; D002454:Cell Differentiation; D004622:Embryo, Mammalian / Q000166:cytology; D005260:Female; D020869:Gene Expression Profiling; D005865:Gestational Age; D007150:Immunohistochemistry; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D008819:Mice, Nude; D046529:Microscopy, Electron, Transmission; D060888:Real-Time Polymerase Chain Reaction; D013724:Teratoma / Q000473:pathology; D015017:Yolk Sac / Q000166:cytology* / Q000648:ultrastructure
chemical_list: D015415:Biomarkers
keywords: Bovine; Embryo; Mesenchymal; Stem cell; Yolk sac