3种分离小鼠腹腔肥大细胞方法的研究

Study on Three Separation Methods of Mast Cells of Mice Abdominal Cavity

In order to get mast cells in mice abdominal cavity,we used three Percoll gradient separation methods to extract mast cells from mice abdominal cavity,and counted cell viability, cell purity and the total number of cells.The 3 mL Percoll gradient separation liquids in the centrifugal tube from top to bottom were divided into 4 layers to analyze the dynamic distribution of mast cells.The results showed that in the method Ⅰ,the cell viability, cell purity and the total number of cells were (83.51±14.00)%，(69.04±11.75)% and（10.60±5.20）×10⁵ /mL,respectively；in the method Ⅱ,the cell viability, cell purity and the total number of cells were (85.71±9.23)%，(87.10±3.93)% and（11.64±5.73）×10⁵ /mL,respectively；in the method Ⅲ,the cell viability, cell purity and the total number of cells were (72.25±24.81)%，(68.34±10.20)% and（8.87±5.18）×10⁵ /mL,respectively.The cell viability and total number of cells in the three groups were not significantly different(P＞0.05).The cell purity of method Ⅱ was extremely significantly higher than that of methods Ⅰ and Ⅲ (Ρ＜0.01). The dynamic distribution of mast cells in the Percoll gradient separation liquid was that the cell viabilities of 3,4 layers were extremely significantly higher than 1 layer (Ρ＜0.01),significantly higher than 2 layer (Ρ＜0.05); the cell purity of 3 layer was extremely significantly higher than 1,4 layers (Ρ＜0.01), significantly higher than 2 layer (Ρ＜0.05); the number of cells of the 3,4 layers were extremely significantly higher than 1,2 layers (Ρ＜0.01). Therefore, adopting the method Ⅱ, layer 3 of extraction, the middle and lower 0.5 mL, which was close to the interface parts in 30%∶ 80% of the Percoll gradient separation liquid, the separation of mast cells of mice peritoneal cavity was best.