Production of senescent cells of Saccharomyces cerevisiae by centrifugal elutriation
Production of senescent cells of Saccharomyces cerevisiae by centrifugal elutriation
复制标题
DOI：
10.1002/yea.320110409
复制
发表时间：
1995-04
期刊：
影响因子：
2.6
通讯作者：
C. Woldringh;K. Fluiter;P. Huls
中科院分区：
文献类型：
--
作者：
C. Woldringh;K. Fluiter;P. Huls
离心洗脱器已被用作婴儿机器，通过将混合代子细胞群体装入腔室，并允许该群体在连续洗涤新生儿子细胞的情况下生长，分裂和老化。洗脱细胞数量的清晰峰至少在十代内可重复获得。在稳态生成时间为95-100 min时，在腔室中生长的亲本细胞继续分裂，在老化过程中循环时间没有变化。洗出的子细胞在前五代的体积从17 μm3的稳态值增加到34 μm3的最大值。正如预期的那样，在同步批次培养中确定的这些大子代的世代时间（130分钟）比稳态子代（240分钟）短，即使来自15代亲本。当一个群体在不清洗较小的子细胞的情况下在腔室中生长时，没有观察到没有芽的子细胞体积增加的迹象。15代亲本群体平均体积为80 μm3，由(i) 71%的细胞有10个以上疤痕，（ii） 13%的细胞有1 - 9个疤痕，（iii） 17%的子细胞组成。通过在洗脱室中不受干扰的生长产生衰老细胞已延长至29代。因此，该方法适用于检查决定出芽酵母寿命的因素。
The centrifugal elutriator has been used as a baby machine by loading the chamber with a population of mixed‐generation daughter cells and allowing this population to grow, divide and age under continuous washing‐out of newborn daughter cells. Clear peaks in the number of elutriated cells were reproducibly obtained for at least ten generations. The parent cells growing in the chamber continued to divide at the steady‐state generation time of 95–100 min, showing no change in cycle time during aging. The washed‐out daughter cells increased in volume during the first five generations from their steady‐state value of 17 μm3 to a maximum of 34 μm3. As to be expected, the generation times of these large daughters, determined in a synchronous batch culture, were shorter (130 min) than that of the steady‐state daughters (240 min), even when derived from 15‐generation parents. No indication for a volume increase of daughter cells without bud was observed when a population was allowed to grow in the chamber without washing‐out the smaller daughter cells. The 15‐generation parent population, recovered from the chamber, had an average volume of 80 μm3 and consisted of: (i) 71% cells with more than ten scars, (ii) 13% cells with one to nine scars, and (iii) 17% daughter cells. The production of senescent cells by undisturbed growth in the elutriator chamber has been prolonged to 29 generations. The method is therefore suitable to examine what factors determine the life span of budding yeast.