Single-Cell Injectoassay for ES Cell Engineering

Abstract

It is important to analyze the intensity dependent action manners of differentiation associated genes in ES cell. For this purpose, single-cell injectoassay was performed by femto-injection method using a single-cell manipulation supporting robot. An Oct3/4 expression vector was introduced into mouse ES cell quantitatively and its effect was evaluated by a cell shape change indicator defined by the perimeter to radius ratio (PR ratio). In the cases of no injection control and the injection control, PR ratio was 7.3-8.0 until 72h. In contrast, when the vector concentration in the injector capillary was 300 ng/micro-L, the rate of differentiation was 33% (2/6). This rate further increased up to 86% (6/7) at 1200 ng/micro-L. These indicate the feasibility of the single-cell injectoassay.