大鼠外周血单个核细胞保存方法研究

通过观察大鼠外周血单个核细胞（PBMCs）的形态，了解不同保存方法和不同保存时期对PBMCs活性和细胞周期分布的影响，建立其短期保存方法．用密度梯度离心法获得大鼠外周血单个核细胞，分别在RPMI1640培养液中4℃保存和冷冻保护剂中－80℃保存，并分别在第1－4天和第7天，14天，21天，28天，35天用台盼蓝染色法测定细胞活力，用流式细胞仪检测细胞周期．结果分离获得的PBMCs成活率为97．69％±1．59％，基本处于G0／G1期．PBMCs置4℃保存4d后，存活率在60．84％±2．75％；置－80℃保存35d后，存活率在78．54％±2．97％，PBMCs基本处于G0／G1期．最后得出－80℃低温保存方法对PBMCs活性和细胞周期检测有一定影响，但其活性率接近80％，是一种简便易行短期保存PBMCs的方法的结论．

Study on Preservation Methods of Peripheral Blood Monouclear Cells of Rat

The gole of study was to explore a simple and effective method of short-term preservation for peripheral blood monouclear cells(PBMCs)of rat by observation morphous of PBMCs.PBMCs got from venous blood of the rat by density gradient centrifugation were preserved in RPMI1640 at 4 ℃ and-80 ℃ refrigerator,respectively.Ttrypan-blue chromophore assay was used to detect the viability and flow cytometry to detect the cell cycle of PBMCs after 1 day,2 days,3 days,4 days,and 7 days,14 days,31 days,28 days,35 days,respectively.The rate of the viability of PBMCs was 97.69 %±1.59 %,and 60.84 %±2.75 % preserved at 4 ℃ for 4 days,and 78.54 %±2.97 % at-80 ℃ for 35 days,respectively.Almost all cells gathered in G0/G1 phase while cell debris added when preservation time prolonged.So preservation of PBMCs at-80 ℃ refrigerator was simple and effective method to keep stability and viability(80 %)of PBMCs.