论文部分内容阅读
本文用荧光细胞分选仪和秋水仙素结合的方法,体外观察小鼠粒单核细胞白血病(MMoL)、粒细胞白血病(L 833-B)细胞的细胞周期和各时相持续时间。体外培养的白血病细胞于指数增殖阶段加入秋水仙素,持续作用12 h,每隔2 h取1组(两瓶)标本,其中1瓶做涂片,计数核分裂细胞数,计算MI;另1瓶用FACS测细胞DNA相对含量,从组方图中计算出每单位时间G_2M期细胞增加率,以及对照组细胞各时相在细胞群中的比例,以便推算细胞Tc及各时相细胞持续时间。用涂片MI方法得到秋水仙素阻断细胞,不同时间核分裂细胞的累积率,其直线回归方程式,MMoL及L 833-B细胞分别为Y=3.10+0.96 X,Y=1.28+0.86 X;Tc分别为104.2和116.3h。Tc太长,可能主要由于涂片时受秋水仙素细胞毒作用后细胞变性,易于破碎,人为降低单位时间MI值。用FACS测定G_2M细胞在秋水仙素作用后其积累率的直线回归方程式,MMoL及L 833 B细胞的分别为Y=26.13+4.01 X,Y=21.04+3.76 X,由此式计算的Tc分别为24.9和26.6 h。这一结果比以往用ARG法所得Tc分别为16和20 h的长些,可能是由于接种的细胞量不同和细胞开始实验时体外培养时间长短不同而引起的,以及秋水仙素对细胞核分裂期除阻断作用外,还有一定杀伤作用和制备标本时对细胞有一定机械性损伤等因素有关。FACS技术用于研究细胞动力学具有快速、简便和避免用放射性核素等优点。
In this study, the cell cycle and the duration of each phase of mouse myelomonocytic leukemia (MMoL) and myeloid leukemia (L 833-B) cells were observed in vitro by the combination of fluorescence cell sorter and colchicine. In vitro, leukemic cells were added with colchicine in the exponential proliferation phase. The cells were continuously treated for 12 h. One group (two bottles) of specimens were taken every 2 h. One of them was smeared and the number of mitotic cells was counted to calculate MI. Using FACS to measure the relative content of cellular DNA, the cell growth rate per unit time in G 2 M phase was calculated from the group diagram, and the proportion of cells in the control group in each cell phase was calculated to calculate the Tc of cells and the duration of cells in each phase. The linear regression equation, MMoL and L 833-B cells were Y = 3.10 + 0.96 X and Y = 1.28 + 0.86 X, respectively, and the cumulative rate of colchicine blocking cells and mitotic cells at different time was obtained by smear MI method. Respectively 104.2 and 116.3h. Tc is too long, may be mainly due to smear cells by colchicine cytotoxicity, easy to break, artificially reduce MI value per unit time. The linear regression equation of the accumulation rate of G_2M cells after colchicine was determined by FACS. The linear regression equations of MMoL and L 833 B cells were Y = 26.13 + 4.01 X and Y = 21.04 + 3.76 X respectively. The Tc calculated by the formula 24.9 and 26.6 h. This result than the previous use of ARG method Tc were 16 and 20 h long, respectively, may be due to the amount of cells inoculated and the cells started experiment in vitro culture time length caused by different, and colchicine on the mitotic In addition to the blocking effect, there are certain killing effects and the preparation of specimens when the cells have some mechanical damage and other factors. FACS technology is used to study the advantages of cellular dynamics such as fast, easy, and avoidance of radionuclides.