Calcium imaging of bioprinted fibers. Active cells are marked out in the micrographs on the left, with the calcium trace in the corresponding color on the right. (A) Day 3 bioprinted fibers showed minimal activity with rapid firing occasionally observed from single myoblasts. (B) Day 7 bioprinted fibers showed a variety of calcium activity, which is commonly observed during myoblast differentiation. (C) By Day 14, active cells had pacemaker activity, characterized by a slower, regular spiking pattern. This is thought to represent a maturation of calcium-handling proteins that can respond to depolarization as well as replenish intracellular stores in time for the next action potential.