Reaggregation of human, chick, and human embryonic brain cells. Factors influencing the formation of a histiotypic unit.

1. Aggregation of embryo human, mouse, and chick brain cells was studied. The optimum age interval of donors from different species was determined. 2. The significance of different dissociation procedures (mild trypsinisation followed by sieving, trypsinisation + DNA digestion, mechanical dissociation in 1 or 2 steps, and Ca2+ chelation by EGTA) for the rate of aggregation was estimated. A significant reduction of aggregation was observed after one step mechanical dissociation. Nonspecific adhesion of cells on DNA molecules was found only during the first stages of aggregation. 3. The curve of aggregation kinetics follows the curve of floculation kinetics. 90% free cells disappear from the medium after 2 h of aggregation and a large number of microaggregates are formed which condense after 20 to 24 h into compact aggregates. The time course of aggregation was similar for all cells dissociated by different means. Small differences in the rate of aggregation, caused by dissociation procedures, were apparent only during the first stages of aggregation. 4. The histiotypic unit formed by aggregation of human, mouse, and chick embryo brain cells exhibits some common and some specific features. During aggregation a multiple structural reconstruction takes place and a limited number of cells are exchanged or sorted out from aggregates into the medium. 5. The structural organisation of aggregates from differently dissociated cells differs in several aspects. This indicates that membrane surface structures are influenced differently by dissociation and behave differently during distinct stages of aggregation.