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      Potential for in vitro mesoderm differentiation of Wharton's jelly cells from ovine umbilical cord isolated in different culture media Translated title: Potencial de diferenciação in vitro de células provenientes da geleia de Wharton de cordões umbilicais ovinos isolados em diferentes meios de cultivo

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          Abstract

          Abstract: The mammalian Wharton's jelly of umbilical cord (WJUC) is a promising source of multipotent cells, providing advantages due to ethical implications, ease of collection and the absence of teratomas in pre-clinical trials. Ovine multipotent cells have already been isolated from various tissues, however there are no reports using umbilical cords in this species. This study aimed to investigate the best medium to transport the umbilical cord, to isolate and maintain ovine WJUC cells and to compare in vitro growth and mesodermal differentiation potential. Eight ovine umbilical cords were obtained during parturition, sectioned and transported in six different media: MEM, low glucose DMEM, M199, RPMI 1640, PBS and saline. For each transportation medium, four culture media were used and the tissue was explanted in 24-well plates and cultured in MEM, low glucose DMEM, M199 and RPMI 1640, all with 10% FBS. Every experiment was conducted with low-passage (P2), investigating MTT viability during four days and adipogenic, chondrogenic and osteogenesis differentiation was induced in vitro. The most effective transport medium (p<0.1) was low glucose DMEM. There was no bacterial or fungal contamination from collection. Cells from Wharton's jelly of ovine umbilical cords collected at natural birth possess fibroblastic morphology and the capacity for in vitro differentiation into adipogenic, chondrogenic and osteogenic cell lines. MTT tests and in vitro differentiation experiments revealed that cell culture medium modulates the behavior of cells and is an important factor for proliferation and maintenance of multipotency. Low glucose DMEM was the most suitable medium for the isolation of cells from Wharton's jelly of ovine umbilical cord.

          Translated abstract

          Resumo: A geleia de Wharton do cordão umbilical (GWCU) de mamíferos é uma fonte promissora de células multipotentes, sendo vantajosa por aspectos éticos, facilidade de coleta e não causar teratomas em ensaios pré-clínicos. Em ovinos, células multipotentes já foram isoladas de vários tecidos, no entanto, não existem relatos do isolamento a partir do cordão umbilical nesta espécie. O objetivo do presente estudo foi investigar o melhor meio para o transporte do cordão umbilical, isolar e manter as células da GWCU ovino em diferentes meios e comparar a proliferação e o potencial de diferenciação mesodermal in vitro. Oito cordões umbilicais foram obtidos, por ocasião do parto natural, seccionados e transportados em seis diferentes meios que consistiram em MEM, DMEM baixa glicose, M199, RPMI 1640, PBS e soro fisiológico. Para cada meio de transporte foram utilizados quatro meios de cultivo, sendo o tecido explantado em placas de 24 poços e cultivados em MEM, DMEM baixa glicose, M199 e RPMI 1640, todos com 10% SFB. Todo o experimento foi realizado em baixa passagem (P2) investigando viabilidade pelo MTT por quatro dias além da indução à diferenciação adipogênica, condrogênica e osteogênica in vitro. O meio de transporte mais efetivo (P<0,10) foi o DMEM baixa glicose. Não houve contaminações bacterianas ou fúngicas decorrentes da coleta. Células oriundas da geleia de Wharton do cordão umbilical ovino colhido por ocasião do parto natural possuem morfologia fibroblastóide e capacidade de diferenciação in vitro nas linhagens adipogênica, condrogênica e osteogênica. Os ensaios de MTT e diferenciação in vitro, revelaram que o meio de cultura celular modula o comportamento destas células, sendo um fator importante tanto para a proliferação como para a manutenção da multipotência, destacando o DMEM baixa glicose como o meio mais adequado para o transporte e isolamento de células da geleia de Wharton do cordão umbilical ovino.

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          Stem cells from foetal adnexa and fluid in domestic animals: an update on their features and clinical application.

          Over the past decade, stem cell research has emerged as an area of major interest for its potential in regenerative medicine applications. This is in constant need of new cell sources to conceive regenerative medicine approaches for diseases that are still without therapy. Scientists drew the attention towards alternative sources such as foetal adnexa and fluid, as these sources possess many advantages: first of all, cells can be extracted from discarded foetal material and it is non-invasive and inexpensive for the patient; secondly, abundant stem cells can be obtained; and finally, these stem cell sources are free from ethical considerations. Cells derived from foetal adnexa and fluid preserve some of the characteristics of the primitive embryonic layers from which they originate. Many studies have demonstrated the differentiation potential in vitro and in vivo towards mesenchymal and non-mesenchymal cell types; in addition, the immune-modulatory properties make these cells a good candidate for allo- and xenotransplantation. Naturally occurring diseases in domestic animals can be more ideal as disease model of human genetic and acquired diseases and could help to define the potential therapeutic use efficiency and safety of stem cells therapies. This review offers an update on the state of the art of characterization of domestic animals' MSCs derived from foetal adnexa and fluid and on the latest findings in pre-clinical or clinical setting of the stem cell populations isolated from these sources.
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            Culture medium modulates the behaviour of human dental pulp-derived cells: Technical Note

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              THE NUTRITION OF ANIMAL TISSUES CULTIVATED IN VITRO

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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Journal
                pvb
                Pesquisa Veterinária Brasileira
                Pesq. Vet. Bras.
                Colégio Brasileiro de Patologia Animal - CBPA. Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA)
                1678-5150
                June 2016
                : 36
                : suppl 1
                : 79-88
                Affiliations
                [1 ] Universidade Estadual do Ceará Brazil
                [2 ]
                [3 ] Universidade da Integração Internacional da Lusofonia Afro-Brasileira Brazil
                [4 ] Universidade Estadual do Ceará Brazil
                Article
                S0100-736X2016001300079
                10.1590/S0100-736X2016001300012
                7018bbd0-eae0-437b-b44d-91fead8b568c

                This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

                History
                Product

                SciELO Brazil

                Self URI (journal page): http://www.scielo.br/scielo.php?script=sci_serial&pid=0100-736X&lng=en
                Categories
                VETERINARY SCIENCES

                General veterinary medicine
                Sheep umbilical cord matrix,multipotent cells,adipogenesis,chondrogenesis,osteogenesis,Matriz do cordão umbilical ovino,células multipotentes,adipogênese,condrogênesse,osteogênese

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