D-Lin-MC3-DMA, an ionizable cationic lipid, is a potent siRNA delivery vehicle.

将DSPC替换为不饱和PCs，并将DLin-MC3-DMA替换为相关脂质DLin-KC2-DMA，可显著提高HeLa细胞体外转染试剂的效能。

White leghorn chicken embryos at stage 24 (30) were removed and dissected in Hanks buffered saline solution (HBSS).?Forelimbs were pooled and treated for 1 h in dispase to remove epithelial ectoderm.?Limb mesenchyme was dissociated, and resuspended in DMEM with 5% FBS and 1% penicillin/streptomycin.?Cells were plated onto 96-well plates at a density of 2x10^5 or 2x10^7 cells/ml and treated with 0-40 μg/ml LNP-pDNA.?Cultures were incubated overnight at 37℃ and resuspended in PBS buffer for expression analysis using a BD LSRII flow cytometer.?Cell survival was assessed in low-density cultures after 24 h of treatment, by counting the adherent cells remaining after PBS washes.?Viability was determined based on average counts of all adherent cells within a single field of view (100x) normalized to cell counts of untreated cultures.