Abstract
We developed a compartmentalized culture system of single embryoid bodies (EBs) utilizing a through-hole on a membrane to induce spatially patterned differentiation. An EB derived from mouse pluripotent stem cells was immobilized on the through-hole. By introducing a stem cell maintenance medium and a differentiation medium into upper and lower culture compartments, respectively, a localized differentiated state was achieved only in the lower part of EB, which is exposed to the medium in the lower compartment. This system may enable us to reconstruct complex tissues and to recapitulate developmental processes using EBs.