Acoustophoresis is promising as a rapid, biocompatible, noncontact cell manipulation method, where cells are arranged along the nodes or antinodes of the acoustic field. Typically, the acoustic field is formed in a resonator, which results in highly symmetric regular patterns. However, arbitrary, nonsymmetrically shaped cell assemblies are necessary to obtain the irregular cellular arrangements found in biological tissues. It is shown that arbitrarily shaped cell patterns can be obtained from the complex acoustic field distribution defined by an acoustic hologram. Attenuation of the sound field induces localized acoustic streaming and the resultant convection flow gently delivers the suspended cells to the image plane where they form the designed pattern. It is shown that the process can be implemented in a biocompatible collagen solution, which can then undergo gelation to immobilize the cell pattern inside the viscoelastic matrix. The patterned cells exhibit F‐actin‐based protrusions, which indicate that the cells grow and thrive within the matrix. Cell viability assays and brightfield imaging after one week confirm cell survival and that the patterns persist. Acoustophoretic cell manipulation by holographic fields thus holds promise for noncontact, long‐range, long‐term cellular pattern formation, with a wide variety of potential applications in tissue engineering and mechanobiology. Acoustic holographic cell manipulation enables complex‐shape cellular pattern formation in a biocompatible hydrogel. Hydrogel encapsulation enables the custom‐designed cell assemblies to be transferred and incubated with good cell viability. This technology holds promise for noncontact, long‐range, long‐term cellular pattern formation, with a wide variety of potential applications in tissue engineering and mechanobiology.