Cytophotometric measurement of the cellular DNA content of [3H]thymidine-labelled spheroids. Demonstration that some non-labelled cells have S and G2 DNA content.

Cell and tissue kinetics Pub Date : 1983-05-01
D C Allison, J M Yuhas, P F Ridolpho, S L Anderson, T S Johnson
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Abstract

Spheroids from the V279-171b and MCa-11 cell lines were incubated continuously for 24 hr in [3H]thymidine for labelling of the outer cells of the viable rim. The spheroids were dispersed into single cells, and the DNA content of photomapped cells was measured by absorption cytophotometry. Autoradiographs were then prepared from which we ascertained cellular labelling. For spheroids of both cell lines, we found a larger proportion of cells with a G0/G1 DNA content among the non-labelled inner spheroid cells than among the labelled outer cells (P less than 0.001). This block of non-labelled spheroid cells in G0/G1 was not a cell cycle perturbation caused by the isotope for the MCa-11 spheroids. Approximately 8% of non labelled MCa-11 spheroid cells had S/G2 DNA content, suggesting that non-cycling cells in spheroids may be blocked in S and G2 as well as in the G0/G1 phase of the cell cycle.

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[3H]胸腺嘧啶标记球体细胞DNA含量的细胞光度测定。证明一些未标记的细胞有S和G2 DNA含量。
将V279-171b和MCa-11细胞系的球体在[3H]胸腺嘧啶中连续孵育24小时,用于标记活膜边缘的外细胞。将这些球体分散到单个细胞中,用吸收细胞光度法测定光疗细胞的DNA含量。然后制备射线自显影片,从中我们确定细胞标记。对于两种细胞系的椭球细胞,我们发现未标记的内椭球细胞中G0/G1 DNA含量的细胞比例大于标记的外椭球细胞(P < 0.001)。G0/G1中未标记的球状细胞块不是由MCa-11球状体的同位素引起的细胞周期扰动。大约8%的未标记的MCa-11球状细胞含有S/G2 DNA,这表明球状细胞中的非循环细胞可能在细胞周期的S和G2以及G0/G1期被阻断。
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