Low intensity pulsed ultrasound (LIPUS) is a therapy used clinically for bone fracture and soft tissue healing LIPUS induces functional alterations in cell behavior and has shown potential for cancer therapy. In order to understand the underlying biophysical mechanisms, the ability to control acoustic exposure parameters and elimination of unwanted effects such as the formation of standing wave patterns, heating, and cavitation, is vital. In this study, we designed an in-vitro focused ultrasound setup (FUS) with repeatable exposure conditions where the cells or spheroids are surrounded by an agar substrate with sufficient absorption to eliminate unwanted reflections and cell culture medium which acts as an effective coolant. The measured maximum temperature rise with the presence of coolant was 1°C and the estimated mechanical index was 1.28. The shear wave propagation induced by acoustic radiation force was monitored and no reflection was observed. We propose an in-vitro FUS cell stimulation setup which can induce non-thermal and non-cavitation biological response with no unwanted artifacts.