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首页> 外文期刊>Cell biology international. >Single-cell heterogeneity in suppression of PC12 differentiation by direct microinjection of a differentiation inhibitor, U0126
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Single-cell heterogeneity in suppression of PC12 differentiation by direct microinjection of a differentiation inhibitor, U0126

机译：通过直接显微注射抑制分化抑制剂，U0126的单细胞异质性。

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Phenotypic and genomic heterogeneity among single cells in a cell population leads to inaccuracy and obscuration in research about mammalian cell differentiation. In order to address the problems regarding bulk analysis on heterogeneous cell populations, it is necessary to accurately regulate and analyze changes in differentiating cells at the single-cell level. To investigate the single-cell changes in PC 12 neuronal differentiation that occur when inhibited by U0126, an inhibitor of mitogen-activated protein kinase kinase (MEK), we directly injected the chemical into individual target cells and analyzed the outcomes (neurite outgrowth) at the single-cell level. As a result, we could accurately regulate the quantity of U0126 being introduced into each target cell, which was previously not possible using the common method of simply adding the inhibitor to the culture medium. It was possible to analyze the inhibitive effect of U0126 even when the injected quantity was lower than the lower limit for inhibition when added to culture medium (0.1 muM, identical to 1.2 x 108 molecules per cell on dish). In particular, injection of 1.5 x 10~7 molecules into each cell resulted in a 59% decrease of the mean total neurite length. Time-course analysis of neurite outgrowth at the single-cell level using fluorescence staining method showed that the changes in neurite length of differentiating PC12 cells were not homogeneous, but were largely variable across individual target cells.
机译：细胞群中单细胞之间的表型和基因组异质性导致哺乳动物细胞分化的研究中的不准确性和遮蔽。为了解决关于异质细胞群体的批量分析的问题，有必要准确调节和分析单细胞水平分化细胞的变化。为了研究PC 12的单细胞变化，在由U0126抑制时发生的PC 12神经元分化，丝裂原激活的蛋白激酶激酶（MEK）的抑制剂，我们将化学物质注入个体靶细胞并分析了结果（神经态过多）单细胞水平。结果，我们可以准确地调节引入每种靶细胞的U0126的量，以前不可能使用简单地将抑制剂添加到培养基中的常见方法。即使当注入的量低于加入培养基时的注射量低于抑制的下限（0.1毫米，在盘上每种细胞上每种细胞的1.2×10 8分子相同的抑制的下限，也可以分析U0126的抑制作用。特别地，注入1.5×10〜7分子到每个电池中，导致平均全神经突长度的59％降低。使用荧光染色方法在单细胞水平下的神经沸石过度的时间过程分析表明，分化PC12细胞的神经突长度的变化不是均匀的，但在各个靶细胞上大大变化。

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