Establishment of a protocol for obtention of neuronal stem cells lineages from the dog olfactory epithelium

Alves, Flávio Ribeiro; Guerra, Ricardo Romão; Fioretto, Emerson Ticona; Delgado, Jeffrey; Júnior, Antônio Augusto Nascimento Machado; Ambrósio, Carlos Eduardo; Kerkis, Irina; Miglino, María Angélica

doi:10.1590/s0100-736x2010000400014

Cited by 6 publications

(8 citation statements)

References 37 publications

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“…Within 5-7 days of explant culture, the spindle-shaped cells were observed migrating out of tissue explant onto the culture dish. Similar observations were observed by Alves et al [10] . The migrated cells possessed typical mesenchymal morphology and continued to proliferate till 7 days.…”

Section: Discussionsupporting

confidence: 92%

“…Primary cell culture was performed according to a previous report [10] . Briefly the mucosal biopsies were dissected under a stereomicroscope to remove cartilage fragments, blood vessels, connective tissues and nonolfactory mucosa.…”

Section: Cell Culturementioning

confidence: 99%

“…However, harvesting such cells directly from the CNS is an invasive procedure with ethical considerations [8] . OM (Olfactory mucosa) of human and dog can easily be obtained from cribriform plate of ethmoidal bone with a non-invasive nasal biopsy [9][10][11][12] . Thus, stem cells derived from canine OM stand as a promising candidate for a source of autologous graft, due to their accessibility [10,12,13] .…”

Section: Introductionmentioning

confidence: 99%

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Köpek Olfaktorik Mukozasindan Olfaktorik Kök Hücrelerin Izolasyonu ve Karakterizasyonu

Altunbaş¹,

Yaprakci²,

Çeli̇k³

2015

Kafkas Univ Vet Fak Derg

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Olfactory stem cells have great potential in the treatment of neurodegenerative diseases and they are good candidates for cell therapy due to the easy accessibility of olfactory mucosa. The main objectives of this study were isolation, proliferation and characterization of olfactory mucosa stem cells that were further differentiated into olfactory neurospheres derived cells. When grown on poly-Dlysine with a serum-free culture medium supplemented with EGF (50 ng/ml) and FGF2 (50 ng/ml), olfactory stem cells gave rise to neurospheres. When grown in serum-containing culture medium newly plated spheres gave rise to olfactory neurosphere derived cells. Gene expression analysis revealed that, OCT4, SOX2, Nanog, Nestin, β-tubulin and NCAM were expressed in olfactory stem cells. While the mRNA expressions of Nanog, Nestin, Oct4, βIII-tubulin and NCAM were downregulated in neurospheres, the mRNA expression of SOX2 upregulated in neurospheres. According to the gene levels of neurospheres generated from olfactory stem cells, beta tubulin and NCAM gene expressions were upregulated, whereas OCT4, Nanog, Sox2 and Nestin mRNA expressions were downregulated in Olfactory neurospheres derived cells. Olfactory mucosa of canine is a suitable alternative source of stem cells and can be applied to cell therapy in neurodegenerative diseases. Keywords: Olfactory stem cell, Canine, Neurosphere, Pluripotent Köpek Olfaktorik Mukozasindan Olfaktorik Kök Hücrelerin Izolasyonu ve Karakterizasyonu ÖzetOlfaktorik kök hücreler nörodejeneratif hastalıkların tedavisinde büyük bir potansiyele sahiptir ve olfaktorik mukozaya kolay erişilebilirliği sayesinde hücre tedavisi için uygun bir adaydır. Bu çalışmanın amacı olfaktorik nörosfer kaynaklı hücrelere kadar farklılaştırılabilen olfaktorik kök hücrelerin izolasyonu, proliferasyonu ve karakterizasyonudur. Olfaktorik kök hücreler EGF (50 ng/ml) ve FGF (50 ng/ml) içeren serumsuz kültür vasatı içerisinde kültüre edildiğinde nörosferleri oluşturdular. Serum içeren kültür vasatında tekrar kültüre edildiklerinde olfaktorik nörosfer kaynaklı hücreleri şekillendirdiler. Gen ekspresyon analizleri OCT4, SOX2, Nanog, Nestin, β-tubulin ve NCAM genlerinin olfaktorik kök hücrelerinde eksprese olduğunu ortaya çıkardı. Nanog, Nestin, Oct4, β tubulin ve NCAM gen ekspresyonları nörosferlerde downregüle olurken, SOX2 geni upregüle oldu. Olfaktorik kök hücrelerin oluşturduğu nörosferlerin gen seviyeleri ile karşılaştırıldığında olfaktorik nörosfer kaynaklı hücrelerde β tubulin ve NCAM gen ekspresyonları upregüle olurken OCT4, Nanog, Sox2 and Nestin mRNA ekspresyonları downregüle oldu. Köpeğin olfaktorik mukozası uygun alternatif bir kök hücre kaynağıdır ve köpek nörodejeneratif hastalıklarında hücre tedavisi için uygulanabilir.

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Section: Discussionsupporting

confidence: 92%

Section: Cell Culturementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Köpek Olfaktorik Mukozasindan Olfaktorik Kök Hücrelerin Izolasyonu ve Karakterizasyonu

Altunbaş¹,

Yaprakci²,

Çeli̇k³

2015

Kafkas Univ Vet Fak Derg

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“…The present study aims to demonstrate the fore mentioned protocol represents an efficient method to rabbit olfactory stem cell characterization and isolation, as it was successfully used to characterize rodents 10,11 and canine 12 OSCs.…”

mentioning

confidence: 99%

Rabbit olfactory stem cells. Isolation protocol and characterization

Ercolin

Roballo²,

Casals³

et al. 2016

Acta Cir. Bras.

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PURPOSE:To describe a new technique for isolation of a mesenchymal stem cells (MSCs) population from the olfactory mucosa in rabbits. METHODS:Olfactory stem cells (OSCs) were retrieved from under the cribriform plate of the Ethmoid bone. Several assays were accomplished to characterize the cell population and attest its viability in vitro. The cells were submitted to flow cytometry with the antibodies CD34, CD45, CD73, CD79, CD90 and CD105 and also they were induced to differentiate in three lineages. Functional evaluation involved analysis of in vitro growth behavior, colony forming unit like fibroblasts (CFU-f) and cryopreservation response.Further transduction with Green Fluorescent Protein (GFP) was also performed. RESULTS:The OSCs showed mesenchymal features, as positive response to CD34, CD73 and CD90 antibodies and plasticity.Additionally, these cells have high proliferated rate, and they could be cultured through many passages and kept the ability to proliferate and differentiate after cryopreservation. The positive response to the transduction signalizes the possibility of cellular tracking in vivo. This is a desirable feature in case those cells are used for pre-clinical trials. CONCLUSION:The cells harvested were mesenchymal stem cells and the technique described is therefore efficient for rabbit olfactory stem cells isolation.

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“…There are numerous animal models, such as rodents 12,15 , swine 16 and canines [17][18][19] that have been used in experiments related to cell therapies. Regarding such animals, outcomes were found in studies about neurodegenerative injuries 12,13 , as well as in attempts to use cell therapy as a way to treat cardiovascular diseases 20 .…”

Section: Introductionmentioning

confidence: 99%

Isolation and characterization of mesenchymal progenitors derived from the bone marrow of goats native from northeastern Brazil

Filho¹,

Neto²,

Carvalho³

et al. 2014

Acta Cir. Bras.

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PURPOSE:To characterize bone marrow progenitors cells grown in vitro, using native goats from northeastern Brazil as animal model. METHODS:Ten northeastern Brazil native goats of both genders were used from the Piauí Federal University Agricultural Science Center's (UFPI) -Goat Farming Sector. Bone marrow aspirates where taken from the tibial ridge and seeded on culture plates for isolation, expansion and Flow Cytometry (expression markers -Oct-3/4, PCNA, Ck-Pan, Vimentina, Nanog). RESULTS:Progenitor cells showed colonies characterized by the presence of cell pellets with fibroblastoid morphology. Cell confluence was taken after 14 days culture and the non-adherent mononuclear cell progressive reduction. After the first passage, 94.36% cell viability was observed, starting from 4.6 x 10 6 cell/mL initially seeded. Cells that went through flow cytometry showed positive expression for Oct-3/4, PCNA, Ck-Pan, Vimentina, and Nanog. CONCLUSIONS:Bone marrow progenitor isolated of native goats from northeastern Brazil showed expression markers also seen in embryonic stem cells (Oct-3/4, Nanog), markers of cell proliferation (PCNA) and markers for mesenchymal cells (Vimentina and Ck-pan), which associated to morphological and culture growth features, suggest the existence of a mesenchymal stem cell (MSC) population in the goat bone marrow stromal cells studied.Key words: Cell Culture Techniques. Cell Differentiation. Stem Cells. Flow Cytometry. Goats. Isolation and characterization of mesenchymal progenitors derived from the bone marrow of goats native from northeastern Brazil

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Establishment of a protocol for obtention of neuronal stem cells lineages from the dog olfactory epithelium

Cited by 6 publications

References 37 publications

Köpek Olfaktorik Mukozasindan Olfaktorik Kök Hücrelerin Izolasyonu ve Karakterizasyonu

Köpek Olfaktorik Mukozasindan Olfaktorik Kök Hücrelerin Izolasyonu ve Karakterizasyonu

Rabbit olfactory stem cells. Isolation protocol and characterization

Isolation and characterization of mesenchymal progenitors derived from the bone marrow of goats native from northeastern Brazil

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