QUANTITATIVE INTRODUCTION OF A CELL DIFFERENTIATION INHIBITING AGENT BY THE USE OF AN AUTOMATED MICROINJECTION SYSTEM
In order to overcome problems in current cell reprogramming methods, such as low efficiency and tumor generation, it is necessary to develop a novel method for quantitative regulation of cell differentiation process and evaluation of its outcomes. By using the automated microinjection apparatus (FUJITSU CI-2500), various agents can be directly injected into cultured cells without the need for intense training or time-consuming operations. This technology enables quantitatively controlled introduction of cell fate-determining factors directly into cultured cells, resulting in precise regulation of the cell differentiation process. In this study, the neuronal differentiation of rat pheochromocytoma cells (PC12) induced by nerve growth factor (NGF) has been quantitatively regulated by introducing ERK inhibitor U0126 into the cells using this automated microinjector. Neurite outgrowth of the differentiated cells has been analyzed on a single-cell basis, using the cell images obtained from the microinjector. Furthermore, comparison of the effect of U0126 injection with the effect of the same inhibitor when added into the culture medium has shown that the use of the microinjector is effective in the quantitative control of the introduction of the substance into each celL As a result, it was shown that the automated microinjection system has the potential to be used for quantitative regulation of cell differentiation and control of the process on a single-cell leveL
cell differentiation cell reprogramming automated microinjector quantitative control
Jihye Chung Natsuko Miura Kouichi Kuroda Mitsuyoshi Ueda
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University,Sakyo-ku, Kyoto 606-8502, Japan
国际会议
重庆
英文
128-129
2011-10-28（万方平台首次上网日期，不代表论文的发表时间）