Hansteen I L, Iversen O H, Refsum S B
Virchows Arch B Cell Pathol Incl Mol Pathol. 1979 Oct;31(2):181-91. doi: 10.1007/BF02889935.
Explants of split mouse ear were incubated in organ culture for up to 48 h, and the cell proliferation was studied by the addition of Thymidine-methyl-3-H (3HTdR) to the medium during different time periods, mainly for the first 14 h of incubation. Cultures were started at 0900, 2130 and 2300. In all cases the labelling index remained stable for 6-8 h, and then increased. The mean grain count, however, was falling and so was the epidermal DNA-specific uptake of 3HTdR. Based on the experimental results, calculations can be made of the flux of cells through S. It is concluded that the increasing LI is not due to inherent diurnal variation in cell proliferation, and is not a sign of real growth but caused instead by a complete block of the cell exit from S, probably combined with periods of an increased entrance rate into S. Other methodological factors, however, may also contribute to the increasing LI. Hence, this system is not suited for the measurement of factors that influence epidermal DNA synthesis.
将分离的小鼠耳部外植体进行器官培养长达48小时,并通过在不同时间段(主要是培养的前14小时)向培养基中添加甲基-3-氢胸腺嘧啶核苷(3HTdR)来研究细胞增殖。培养在09:00、21:30和23:00开始。在所有情况下,标记指数在6 - 8小时内保持稳定,然后增加。然而,平均颗粒计数在下降,3HTdR的表皮DNA特异性摄取也在下降。根据实验结果,可以计算出细胞通过S期的通量。得出的结论是,标记指数的增加不是由于细胞增殖固有的昼夜变化,也不是真正生长的标志,而是由细胞从S期完全阻滞导致的,可能还伴有进入S期的速率增加的时期。然而,其他方法学因素也可能导致标记指数增加。因此,该系统不适合用于测量影响表皮DNA合成的因素。