Aggregation enhances catecholamine secretion in cultured cells

Transplanted cells and tissues have potential uses in the treatment of genetic, geriatric, and metabolic disorders, but optimal conditions for transplantation are not yet known. In this report, PC12 cells were aggregated in rotary and microgravity culture, using serum-free or serum-supplemented medium, and using a multifunctional polymer-peptide aggregation factor. Aggregates and single cells were then encapsulated and cultured within agarose gels, and the dopamine secretion in response to a depolarization buffer was measured using high-performance liquid chromatography combined with electrochemical detection (HPLC-ECD). On a per-cell basis, aggregated cells secreted higher levels of dopamine than did single cells. The size of the aggregates was also a factor in catecholamine secretion; dopamine release from the larger aggregates formed in rotary culture was observed to increase at a faster rate, then achieve a plateau level at an earlier time than did the smaller aggregates. Cells aggregated in microgravity culture exhibited a markedly different behavior, lacking the rapid rise in dopamine secretion characteristic of the rotary-aggregates cells: on a per-cell basis, the dopamine secretion remained at a level corresponding to the plateau level expressed by the rotary-aggregates cells. Dopamine secretion in aggregates may be enhanced by the increase in number of cell-cell contacts, as occurs during high-density culture of PC12 cells. These results provide further evidence that cell-cell contact regulates the behavior of differentiated cells, and therefore is important in tissue engineering.