. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. Figure 1. The sequential processes of Mytiliis califomianus hemocyte aggregation in vitro, (a) The naive free cell stage: these fresh hemocytes are approximately spherical in outline, (b) The initial stage of cell ag- gregation and adhesion: Within one minute, all of the cells form spikes, and some cells form small aggregates (cell number < 10). (c) Medium sized clumps: when these small aggregates collide, they can form bigger clumps. Each medium sized clump may be composed of about one hundred cells. Bars = 30 ^ Kigur
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. Figure 1. The sequential processes of Mytiliis califomianus hemocyte aggregation in vitro, (a) The naive free cell stage: these fresh hemocytes are approximately spherical in outline, (b) The initial stage of cell ag- gregation and adhesion: Within one minute, all of the cells form spikes, and some cells form small aggregates (cell number < 10). (c) Medium sized clumps: when these small aggregates collide, they can form bigger clumps. Each medium sized clump may be composed of about one hundred cells. Bars = 30 ^ Kigure 2. .\/iv//».s califomianus hemocyte spreading and migration in viiro (a) Part of a large clump: each large aggregate like this may contain more than ten thousand cells. These cohere tightly together, and the individual cells are difficult to distinguish. Bar = 30 ^m. (b) Early stage of hemocyte migration: cells at the edge begin to move out of the aggregate about 3 minutes after the aggregate settles down. Bar = 30 nm. (c) Late stage of hemocyte migration: cells have radially migrated from the aggregate after 1(1 min in vitro Bar = 50 pm. were distinguishable and could be easily dispersed from the aggregates by pipetting. In contrast, clumps that formed during 10 or more minutes of shaking often contained more than ten thou- sand cells (Fig. 2a). This kind of clump was resistant to mechanical dispersion by pipetting or vortexing. When cells or aggregates were allowed to settle down onto glass or plastic, they attached, flattened, and developed pseu- dopodia. During subsequent incubation, cells located at the surface of the clump and in contact with the substra- tum adhered to the substratum and migrated radially out of the clump (Fig. 2b, c). The attached and spread cells moved by amoeboid locomotion. The cells possess a strong ability to adhere, spread, and migrate on foreign surfaces. Only on agarose substrata were such kinds of cell behaviors inhibited (Table I). Assay of
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