We present a microfluidic chip for cell handling applications fabricated in polydimethylsiloxane (PDMS). The chip was designed to accumulate cells inside a reaction chamber where they can be manipulated under attendance of a liquid reagent (PEG–polyethylene glycol) for cell fusion. To enable the cell handling a fluid management system was developed which includes pneumatically actuated microfluidic valves integrated into the chip. Various valve geometries were simulated in ANSYS and characterized in a series of tests to identify a feasible valve concept. The main criterion of the valve development was a reliable closing characteristic. This characteristic was simulated in ANSYS and was verified experimentally. Both, the microfluidic layer and the pneumatic layer, were realized by means of soft-lithographic techniques and bonded using a plasma enhanced bonding process. First test series showed the eligibility of the applied valve concept and of the microfluidic chip for cell handling and for cell fusion.