A Method for Generation of Bone Marrow-Derived Macrophages from Cryopreserved Mouse Bone Marrow Cells

The broad use of transgenic and gene-targeted mice has established bone marrow-derived macrophages (BMDM) as important mammalian host cells for investigation of the macrophages biology. Over the last decade, extensive research has been done to determine how to freeze and store viable hematopoietic human cells; however, there is no information regarding generation of BMDM from frozen murine bone marrow (BM) cells. Here, we establish a highly efficient protocol to freeze murine BM cells and further generate BMDM. Cryopreserved murine BM cells maintain their potential for BMDM differentiation for more than 6 years. We compared BMDM obtained from fresh and frozen BM cells and found that both are similarly able to trigger the expression of CD80 and CD86 in response to LPS or infection with the intracellular bacteria Legionella pneumophila. Additionally, BMDM obtained from fresh or frozen BM cells equally restrict or support the intracellular multiplication of pathogens such as L. pneumophila and the protozoan parasite Leishmania (L.) amazonensis. Although further investigation are required to support the use of the method for generation of dendritic cells, preliminary experiments indicate that bone marrow-derived dendritic cells can also be generated from cryopreserved BM cells. Overall, the method described and validated herein represents a technical advance as it allows ready and easy generation of BMDM from a stock of frozen BM cells

Análise comparativa da função respiratória de indivíduos hígidos em solo e na água

A mensuração da função respiratória oferece informações essenciais para caracterizar anormalidades pulmonares. A pressão hidrostática da água atua no tórax submerso de diversas formas, causando alterações no sistema respiratório. O objetivo deste estudo foi analisar comparativamente variáveis que avaliam a função respiratória - volume minuto (Vmin), volume corrente (Vc), capacidade vital (Cvital) e frequência respiratória (FR) - de voluntárias no solo e com o tórax submerso em piscina terapêutica aquecida. A função respiratória de 30 voluntárias saudáveis (20,9±2,1 anos; 1,64±0,07 m; 58,8±9,2 kg; índice de massa corporal 21,78±2,63 kg/m²) foi avaliada por meio de ventilômetro em solo e aos 1 e 20 minutos de imersão, com água ao nível dos ombros, em posição sentada. Após 20 minutos de imersão, foi registrado aumento estatisticamente significativo no Vmin (p=0,015) e Vc (p=0,027); e uma redução estatisticamente significativa (p=0,016) na Cvital 1 minuto após imersão, em relação aos valores obtidos em solo. O maior tempo de imersão alterou assim os valores obtidos em solo, com exceção da Cvital, que sofreu alteração significativa desde o primeiro minuto de imersão. Não foram encontradas diferenças significativas entre os valores obtidos após 1 e 20 minutos na água. O estudo permite concluir que a imersão do tórax em piscina aquecida provocou aumento no Vmin e Vc e diminuição na Cvital de voluntárias saudáveis.Measuring respiratory function provides essential information to assess pulmonary changes. Effects of water hydrostatic pressure on the submerged chest cause changes in the respiratory system. The purpose here was to compare respiratory function variables - minute volume (MV), tidal volume (TV), vital capacity (Vitalc), and respiratory rate (RR) - on the ground and with chest submerged in water. Respiratory function of 30 healthy female volunteers (mean age 20.93 ± 2.11; weight 58.8±9.2 kg; body mass index 21.78±2.63 kg/m²) was assessed by spirometry on the ground, and 1 and 20 minutes after immersion in warm water at shoulder level in the sitting position. As compared to ground levels, statistically significant increases were found in MV (p=0.015) and TV (p=0.027) 20 minutes after immersion, as well as a significant decrease (p=0.016) in Vitalc one minute after immersion. Longer time immersion has thus altered values obtained on ground, except for Vitalc, which showed significant reduction on the first minute after chest immersion. Comparison between variable values obtained 1 and 20 minutes in water showed no significant difference. It may thus be said that chest submersion in warm water caused an increase in MV and VT and a decrease in Vitalc of healthy subjects