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

Pesquisa Veterinária Brasileira

Flávio R. Alves²  Ricardo R. Guerra¹  Emerson T. Fioretto¹  Juliana C. Delgado¹  Antônio A. N. Machado Júnior²  Carlos E. Ambrósio¹  I. Kerkis¹  Maria A. Miglino¹ 

[1] ,Universidade Federal do Piauí Curso de Medicina Veterinária Disciplina Diagnóstico por ImagemBom Jesus PI ,Brazil

关键词: Olfactory epithelium;    canine cell culture;    animal stem cell;    Epitélio olfatório;    cultivo de células caninas;    células-tronco animais;   

DOI  :  10.1590/S0100-736X2010000400014

来源: SciELO

【 摘 要 】

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.

【 授权许可】

CC BY-NC   
 All the contents of this journal, except where otherwise noted, is licensed under a Creative Commons Attribution License