Abstract Good cell culture practices are a set of technical and management tools recommended for application in research and service laboratories to guarantee the traceability and reproducibility of in vitro experiments. However, most research laboratories do not have a structured quality assurance system and have difficulties organizing their workflows or even priorities in implementing acceptable laboratory practices. In this study, we applied management and quality assurance tools to define the steps necessary to implement acceptable laboratory practices in the multiuser laboratory of cell culture and establish a cell bank at the Carlos Chagas Institute FIOCRUZ-PR. Our team applied the 5W2H and 5S tools for initial diagnosis and established an action plan to implement and manage the laboratory over two years. Thereby, we defined the scope of laboratory activities, including the demand for establishing a cell bank, the supply of cell lines to internal users, user training, and quality control tests. We also mapped the main activities, establishing their flows and all the necessary documentation to ensure traceability and reproducibility. Additionally, the laboratory was organized in compliance with the 5S principles. In conclusion, using management tools, such as the 5W2H and 5S methods, is a simple and affordable method to organize and manage a cell culture laboratory and can be applied to other research laboratories.

RESUMO Objetivo: Avaliar a capacidade do eluato proveniente da membrana de plasma rico em fibrina (PRF) de induzir proliferação e diferenciação das células-tronco humanas isoladas de tecido adiposo (CTDAh) em condrócitos. Método: As membranas de PRF foram obtidas por centrifugação de sangue periférico de dois indivíduos saudáveis, cortadas, colocadas em poços de placa de cultivo por 48 h para obtenção do eluato de fibrina. As CTDAh foram isoladas do tecido adiposo por digestão com solução de colagenase e expandidas in vitro. As células foram expandidas e tratadas com meio de cultivo DMEM-F12, meio comercial para diferenciação condrocítica, e eluato de fibrina durante três dias e marcadas com BrdU para avaliação quantitativa da proliferação celular com o uso do sistema de imagens High-Content Operetta®. Para o ensaio de diferenciação condrogênica, as CTDAh foram cultivadas em micromassa por 21 dias e coradas com azul de toluidina e agrecana para avaliação qualitativa em microscópio óptico. As avaliações estatísticas foram feitas por meio dos testes Anova e Tukey. Resultados: Houve uma maior proliferação das células tratadas com o eluato de fibrina comparativamente com os outros dois tratamentos, nos quais o teste Anova apontou significância (p < 0,001). A diferenciação em condrócitos foi visualizada pela presença de mucopolissacarídeos na matriz das células tratadas com meio de diferenciação ou eluato e marcação positiva para agrecana. Conclusões: O tratamento com o eluato da membrana de fibrina estimulou a proliferação celular e induziu a diferenciação das células-tronco em condrócitos, o que sugere uma potencial aplicação da membrana de PRF nas terapias de regeneração de cartilagem hialina.

ABSTRACT Objective: To evaluate the ability of the eluate from fibrin-rich plasma (FRP) membrane to induce proliferation and differentiation of isolated human adipose-derived stem cells (ASCs) into chondrocytes. Method: FRP membranes were obtained by centrifugation of peripheral blood from two healthy donors, cut, and maintained in culture plate wells for 48 h to prepare the fibrin eluate. The SCATh were isolated from adipose tissue by collagenase digestion solution, and expanded in vitro. Cells were expanded and treated with DMEM-F12 culture, a commercial media for chondrogenic differentiation, and eluate from FRP membrane for three days, and labeled with BrdU for quantitative assessment of cell proliferation using the High-Content Operetta® imaging system. For the chondrogenic differentiation assay, the SCATh were grown in micromass for 21 days and stained with toluidine blue and aggrecan for qualitative evaluation by light microscopy. The statistical analysis was performed using ANOVA and Tukey's test. Results: There was a greater proliferation of cells treated with the eluate from FRP membrane compared to the other two treatments, where the ANOVA test showed significance (p < 0.001). The differentiation into chondrocytes was visualized by the presence of mucopolysaccharide in the matrix of the cells marked in blue toluidine and aggrecan. Conclusions: Treatment with eluate from FRP membrane stimulated cell proliferation and induced differentiation of the stem cells into chondrocytes, suggesting a potential application of FRP membranes in hyaline cartilage regeneration therapies.

Com o objetivo de avaliar o crescimento e o acúmulo de nutrientes por híbridos de melão desenvolveram-se experimentos em área comercial no município de Baraúna-RN, e na estação experimental de Bebedouro (Petrolina-PE) da Embrapa Semiárido. Em Baraúna-RN, o experimento foi desenvolvido de setembro a novembro de 2010, e em Petrolina-PE, de agosto a outubro de 2010. O delineamento experimental foi em blocos casualizados completos, com quatro repetições, em esquema fatorial 2 x 5. Os tratamentos resultaram da combinação de dois híbridos de melão (Iracema e Gran Prix) e cinco épocas de coletas de plantas (15; 25; 35; 45 e 55 dias após o transplante, DAT). Para se fazer um programa de adubação na cultura do meloeiro, tanto do grupo Amarelo como do grupo Pele-de-Sapo, observou-se um crescimento lento no início, maior incremento da parte aérea de 35 a 45 DAT e maior exigência da planta para o estádio de frutificação de 35 a 45 DAT para 'Iracema' e 45 a 55 DAT para 'Gran Prix' Quanto ao acúmulo, para os dois materiais e nos dois locais, observou-se a seguinte ordem: K>N>P>Ca>Mg.

Aiming to evaluate the growth and nutrient accumulation by hybrid melon experiments were developed in commercial area in the city of Baraúna - RN and in the experimental station Embrapa Semi-Arid (Petrolina - PE). In Baraúna - RN, the experiment was conducted from September to November 2010 and in Petrolina - PE from August to October 2010. The experimental design was a completely randomized design with four replications in a factorial 2 x 5. Treatments resulted from the combination of two melon hybrids (Iracema and Gran Prix) and five periods of the gathering of samples plants (15, 25, 35, 45 and 55 days after transplanting 'DAT'). To make a fertilization program in the melon, such as the Yellow Group as the Group of Skin Frog, there was a slow growth in the beginning, the greater increase in aerial parts was from 35 to 45 DAT, and the requirement for the fruiting stage from 35 to 45 DAT for Iracema and 45 to 55 DAT for Grand Prix; was observed the following order: K > N > P > Ca > Mg for the accumulation of nutrients in the two samples of the two cities.