Computational protocol: Maturation of dendritic cells in vitro and immunological enhancement of mice in vivo by pachyman- and/or OVA-encapsulated poly(d,l-lactic acid) nanospheres

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Protocol publication

[…] At the end of the stimulation, the maturation status of BMDCs was assessed. In brief, the co-cultured and undisposed BMDCs were gathered, rinsed twice with PBS, and three fold-labeled for half an hour at 4°C, protected from light, with APC-anti-mouse CD11c, FITC-anti-mouse CD80, and PE-anti-mouse CD86 monoclonal antibodies (BD Biosciences) as DC maturation markers. The corresponding isotype control antibodies were used as controls. After thorough rinsing by washing twice with PBS, expression levels of associated surface molecules in labeled BMDCs were determined by flow cytometry in a FACS Calibur cytometer (BD Biosciences). The data were analyzed using FlowJo software (version 7.6). [...] All experiments were performed in triplicate, and data are expressed as mean ± standard error of the mean (SEM). Statistical differences between groups were determined by one-way analysis of variance (ANOVA) followed by Duncan’s multiple range test with IBM SPSS Statistics 20.0 for analysis. Differences regarded as statistically significant between the groups are presented as follows: *p<0.05, **p<0.01, and ***p<0.001 (compared with the PHY group); and #p<0.05, ##p<0.01, and ###p<0.001 (compared with the PLA group). […]

Pipeline specifications

Software tools FlowJo, SPSS
Applications Miscellaneous, Flow cytometry
Organisms Mus musculus