The objective of this study was to describe guidelines for the isolation of bone marrow mononuclear cells from rabbits, followed by cell purification by negative depletion with CD45 monoclonal antibody, and further expansion in MesenCult ® medium. Ten adult male New Zealand White rabbits, age average of 1.0±0.2 years and weighting 3.5±0.24kg, were used to obtain a standardized method. The mononuclear cells of the bone marrow were isolated with Ficoll-paque ® density gradient centrifugation, and the cell purification and acquisition was completed by negative depletion with CD45 monoclonal antibody in immunomagnetic base. The cell population obtained was expanded in MesenCult ® medium. Through isolation with Ficoll-paque ® density gradient was possible to obtain an average yield of 7.31x10 6 cells/mL. After purification and acquisiton of potential mesenchymal stem cells by the immunomagnetic base, there was a yield decrease to 2.28x10 6 cells/mL; however the expansion process was increased in cell culture. The results indicated that cells obtained from the mononuclear fraction of bone marrow and cultivated in vitro were capable to generate adherent cells 24 hours after culture, with predominance of fibroblastoid cells suggestive of mesenchymal stem cells. It can be concluded that mesenchymal stem cells can be achieved with purified rabbit bone marrow mononuclear cells through the immunomagnetic method, as the MesenCult ® medium provides a suitable environment for a quick in vitro expansion, and the number of passages exerts negative influence on the morphological characteristics.INDEX TERMS: Mesenchymal stem cells, lagomorph, immunomagnetic separation.

Pesq. Vet. Bras. 33(9):1076-1080, setembro 2013 1076 RESUMO.-[Resíduos de sanitizantes em vacas de leite após passagem por pedilúvio.] O sulfato de cobre e o hipoclorito de sódio são empregados na prevenção e tratamento das enfermidades digitais dos bovinos, mas os valores residuais desses elementos foram pouco estudados. Neste estudo, avaliou-se a presença de resíduos de cobre e cloretos totais no leite de vacas saudáveis após passagens dos animais em pedilúvio contendo soluções formuladas com estas substâncias e estimou-se os custos das soluções. Utilizou-se 14 vacas saudáveis distribuí-das em dois grupos (GI e GII) de sete animais cada. Em GI, empregou-se solução de hipoclorito de sódio a 1% e, em GII, sulfato de cobre a 5%. As amostras de leite foram colhidas antes da passagem pelo pedilúvio (M0), após 24 (M1), 48 (M2) e 72 (M3) horas, além de 15 dias (M15) subsequentes à última passagem. Na análise estatística, a comparação entre momentos dentro de cada grupo foi realizada com teste de Friedman, seguido pelo teste de Dunn's (p<0,05). Concluiu-se que os valores de cloretos totais e de cobre no leite de bovinos saudáveis, após passagens diárias dos animais em pedilúvio por um período de sete dias, apresentaram algumas variações consideradas insuficientes para provocarem danos à saúde humana Copper sulfate and sodium hypochlorite are used in footbath solutions for the prevention and treatment of bovine digital diseases; however, data on the residues of such elements in milk are sparse in Brazil. This study evaluated the cost of applying the footbath treatment and the total amount of copper and chlorite residues in the milk of healthy cows after they had passed through these footbath solutions. Two groups of 7 cows each (GI and GII) were studied. In the case of GI, 1% sodium hypochlorite was used and for GII 5% copper sulfate was employed in the footbath. The milk samples were collected before the 7-day footbath treatment period (M0) and 24 h (M1), 48 h (M2), 72 h (M3) and 15 days (M15) after the last footbath. Statistical analysis to compare the different samples within each group was carried out by applying Friedman's test, followed by Dunn's test (p<0.05). It was concluded that the amount of total chlorites and copper in the milk of healthy cattle after routine daily footbaths for a period of 7 days presented some variations. However, the concentrations observed were considered insufficient to represent a risk to human health. The cost of the footbath solutions was found to be reasonable.