We developed a method for noninvasively harvesting cells, cell sheets, and spheroids from a culture surface along with the electrochemical desorption of a self-assembled monolayer (SAM). Cells were attached on a gold culture surface on which Arg-Gly-Asp (RGD) containing peptides were covalently bonded via the SAM of an alkanethiol. The application of ?1.0 V (vs Ag/AgCl) caused the reductive desorption of the SAM, resulting in the detachment of the cells. By this method, more than 90% of the cells were detached within 5 min. Two-dimensional (2D) cell sheets could also be detached from the gold surface in the same manner. The detached cell sheets consisted of viable cells that could easily attach to other cell sheets in succession and form a multilayered cell sheet. Moreover, by combining this approach with photolithography and micro-contact printing (μCP) technologies, we fabricated a chip system in which hepatocytes formed spheroids of a uniform diameter at the density of 280/cm2 and could be harvested by the application of a negative potential. This cell manipulation technology could potentially be a useful tool for the fabrication and assembly of building blocks such as cell sheets and spheroids for regenerative medicine and tissue engineering applications.