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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

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

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Author(s):
Alves, Flavio R. [1] ; Guerra, Ricardo R. [2] ; Fioretto, Emerson T. [3] ; Delgado, Juliana C. [4] ; Machado Junior, Antonio A. N. [1] ; Ambrosio, Carlos E. [5] ; Kerkis, I. [6] ; Miglino, Maria A. [4]
Total Authors: 8
Affiliation:
[1] UFPI, Curso Med Vet, Disciplina Diagnost Imagem, BR-64900000 Bom Jesus, PI - Brazil
[2] Univ Fed Paraiba, Ctr Ciencias Agr, Dept Ciencias Vet, BR-58397000 Areia, PB - Brazil
[3] Univ Fed Sergipe, Ctr Ciencias Biol & Saude, Dept Morfol, Lab Biol Celular & Estrutural, BR-49100000 Sao Cristovao, SE - Brazil
[4] Univ Sao Paulo, Dept Cirurgia, FMVZ, BR-05508270 Sao Paulo - Brazil
[5] Univ Sao Paulo, FZEA, Dept Ciencias Basicas, BR-13635270 Pirassununga, SP - Brazil
[6] Inst Butantan, Genet Lab, BR-05503900 Sao Paulo - Brazil
Total Affiliations: 6
Document type: Journal article
Source: Pesquisa Veterinária Brasileira; v. 30, n. 4, p. 363-372, APR 2010.
Web of Science Citations: 5
Abstract

A morphological and cell culture study from nasal mucosa of dogs was performed in order to establish a protocol to obtain a cell population committed to neuronal lineage, as a proposal for the treatment of traumatic and degenerative lesions in these animals, so that in the future these results could be applied to the human species. Twelve mongrel dogs of 60-day aged pregnancy were collected from urban pound dogs in São Paulo. Tissue from cribriform ethmoidal lamina of the fetuses was collected at necropsy under sterile conditions around 1h to 2h postmortem by uterine sections and sections from the fetal regions described above. Isolated cells of this tissue were added in DMEM/F-12 medium under standard conditions of incubation (5% CO², >37ºC). Cell culture based on isolated cells from biopsies of the olfactory epithelium showed rapid growth when cultured for 24 hours, showing phase-bright sphere cells found floating around the fragments, attached on culture flasks. After 20 days, a specific type of cells, predominantly ellipsoids or fusiform cells was characterized in vitro. The indirect immunofluorescence examination showed cells expressing markers of neuronal precursors (GFAP, neurofilament, oligodendrocyte, and III â-tubulin). The cell proliferation index showed Ki67 immunostaining with a trend to label cell groups throughout the apical region, while PCNA immunostaining label predominantly cell groups lying above the basal lamina. The transmission electron microscopy from the olfactory epithelium of dogs revealed cells with electron-dense cytoplasm and preserving the same distribution as those of positive cell staining for PCNA. Metabolic activity was confirmed by presence of euchromatin in the greatest part of cells. All these aspects give subsidies to support the hypothesis about resident progenitor cells among the basal cells of the olfactory epithelium, committed to renewal of these cell populations, especially neurons. (AU)