Brás Susana, França Ângela, Cerca Nuno
Centre of Biological Engineering (CEB), Laboratory of Research in Biofilms Rosário Oliveira (LIBRO), University of Minho, Braga, Portugal.
PeerJ. 2020 Jun 15;8:e9295. doi: 10.7717/peerj.9295. eCollection 2020.
Human blood is often used as an ex vivo model to mimic the environment encountered by pathogens inside the host. A significant variety of experimental conditions has been reported. However, optimization strategies are often not described. This study aimed to evaluate key parameters that are expected to influence gene expression when using human blood ex vivo models. Our data confirmed that blood antimicrobial activity was dependent on initial bacterial concentration. Furthermore, blood degradation over time resulted in lower antimicrobial activity, with a 2% loss of leukocytes viability correlating with a 5-fold loss of antimicrobial activity against . We further demonstrated that the volume of human blood could be reduced to as little as 0.18 mL without affecting the stability of gene expression of the tested genes. Overall, the data described herein highlight experimental parameters that should be considered when using a human blood ex vivo model for gene expression analysis.
人体血液常被用作体外模型，以模拟病原体在宿主体内所遇到的环境。已有大量不同的实验条件被报道。然而，优化策略往往未被描述。本研究旨在评估在使用人体血液体外模型时，预期会影响基因表达的关键参数。我们的数据证实，血液的抗菌活性取决于初始细菌浓度。此外，随着时间推移血液降解导致抗菌活性降低，白细胞活力损失2%与针对……的抗菌活性损失5倍相关。我们进一步证明，人体血液体积可减少至低至0.18 mL，而不影响所测试基因的基因表达稳定性。总体而言，本文所述数据突出了在使用人体血液体外模型进行基因表达分析时应考虑的实验参数。
