Cytological studies on cellular antigen-antibody reaction in tissue culture. Observations by means of a newly devised culture chamber under the phase contrast microscope
Mie Med Jour 8(2): 211-230
A newly devised culture chamber is described, which permits continuous observation of cultivated cells under the phase contrast microscope and during the exchange of supernatant fluid medium. By means of the chamber, tissue-monocytes from the peritoneal fluid of sensitized rabbits were cultivated and observed before and after contact with the antigen. Some attempts were made to cultivate tissue-monocytes having an extended form and clear cytoplasm so as to observe in detail the structural changes. The use of diluted supernatant fluid medium with ultrafiltrated horse serum served to achieve this purpose. Shortly after contact with a homologous antigen, most of the tissue-monocytes showed primarily retraction of the processes, marked swelling of the cytoplasm, formation of fibrous meshwork in the swollen cytoplasm, decolorization of Neutral Red and Janus Green granules and diffuse staining of the cytoplasm and nucleus. Structural change mitochondria appeared later than in the processes and the ground cytoplasm. Golgi bodies of the tissue-monocytes were sometimes observed in the nuclear cavity, which showed a reticular structure composed of three parts; the internal clear space, the external reticular structure, and the periexternal clear zone. The structural changes in cellular antigen-antibody reaction were also of secondary nature. Some observations of the tissue-monocytes were described, which were at first refractory to the antigen but showed structural changes after 30 minutes. The average proportion of these cells was approximately 5%. Prolonged observations from 1.5 to 24 hours did not show progressive changes in the injured cells. Tissue-monocytes upon contact with a heterologous antigen did not show the characteristic changes described above.