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兔中耳上皮细胞体外生长与合成的定量分析

Quantification of in vitro growth and synthesis in rabbit middle ear epithelial cells.

作者信息

Schousboe L P, Blegvad S, Ovesen T

机构信息

ENT Department, Aarhus University Hospital, Denmark.

出版信息

Eur Arch Otorhinolaryngol. 1996;253(7):395-400. doi: 10.1007/BF00168490.

Abstract

The purpose of the present study was a quantitative description of middle ear epithelial cell reactions in a standardized model system under controlled conditions. The model is based on a newly developed culture method for rabbit middle ear epithelial cells. The growth pattern of identical subcultures was determined by measurement of total cell protein, DNA synthesis and DNA amount during a 15-day period. The secretory product was quantified by means of 3H-glucosamine incorporation. Epithelial cells from the auditory meatus were examined by identical methods. The results proved the method to be reproducible regarding all parameters. The DNA amount and total cell protein increased throughout the period, whereas DNA synthesis remained constant. Secretion increased linearly for 9 days and then diminished. All parameters were without significant changes when related to DNA amount, reflecting a uniform cellular activity throughout the period. The DNA amount was the most precise parameter with a mean variation coefficient of 6.0% (SD 3.1%). With the baseline values found to be reproducible for the parameters chosen, our model system is considered to be usable for quantifying various factors influencing the proliferation and activity of middle ear epithelium.

摘要

本研究的目的是在可控条件下,对标准化模型系统中的中耳上皮细胞反应进行定量描述。该模型基于一种新开发的兔中耳上皮细胞培养方法。通过在15天内测量总细胞蛋白、DNA合成和DNA量,确定相同传代培养物的生长模式。分泌产物通过3H-葡萄糖胺掺入法进行定量。采用相同方法检查外耳道上皮细胞。结果证明该方法在所有参数方面均可重复。在此期间,DNA量和总细胞蛋白均增加,而DNA合成保持恒定。分泌在9天内呈线性增加,然后减少。与DNA量相关时,所有参数均无显著变化,这反映了整个期间细胞活性的一致性。DNA量是最精确的参数,平均变异系数为6.0%(标准差3.1%)。由于所选参数的基线值具有可重复性,我们的模型系统被认为可用于量化影响中耳上皮增殖和活性的各种因素。

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