Cerebral neurons, skeletal myoblasts, and cardiac muscle cells cultured on macroporous beads.

Abstract

Biodegradable macroporous spherical microcarriers (MCs) offer a suitable substrate for adhesion, growth, and differentiation of cerebral neurons, skeletal myoblasts, and cardiac cells. The cavernous structure of these gelatin beads supplies a large surface and a tridimensional habitat for cell propagation. Within hours from their seeding, all three cell types became firmly attached to the MCs, forming cell-MC aggregates, which remained floating in the medium. Neuronal aggregates were composed mainly of single or groups of perikarya, and their sprouted nerve processes formed a ramified network. In skeletal muscle aggregates, fusion of myoblasts into myotubes occurred within 5 days. The myotubes, arranged in bundles having the same orientation, became striated and the whole aggregate, or parts of it, contracted spontaneously. Cardiac cells divided in the aggregate to form one or more layers of flat cells, which exhibited single microvilli. The cells were interconnected at uneven intervals and the whole aggregate contracted actively. Morphological and biochemical analysis of sampled aggregates ensured that cells reached the proper stage of maturation and could be used for either physiological and pharmacological studies or implantation into injured or dystrophic tissue.