This paper describes the fabrication and the evaluation of a reusable Cell Culturing Device, designed for a single cell and cellular networks analysis. This is the first success of combination of Microcontact Printing (mCP) and Vacuum Device. This combination has following advantages; (1) cells stay within the micropatterns for long enough duration to achieve local activation of cells or cellular networks (more than 24 h), (2) the displacement distance of laminar flow from the interface of two fluids in Cell Culturing Device keeps smaller than the diameter of a cell, (3) all components of our device except micropatterned substrate are reusable for further analyses. The success of the combination of above techniques provides a controllable environment for the local activation of a single cell and cellular networks. Our device allows to exhibit the different responses induced with the various conditions in a single observation sight at exactly the same time point.