Direct conversion of precursors of PFCs into active PFCs in vitro, without prior cell division.
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Biomedical subjects
Publications and source records attributed to J Marbrook.
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The BSA density-gradient technique for separating mouse spleen cells into partially purified populations has been used to compare the responsiveness of such populations to SRBC using in vivo and in vitro techniques. Two major populations were distinguished, one of which responded very well in vivo with an exponential dose response and poorly in vitro (fraction 3), and another which responded in vivo and in vitro with a linear dose response (fraction 2). A light density, radiation-resistant component was identified which markedly stimulated the response of fraction 3 in vitro, and a density gradient profile was obtained for this cell which did not correspond with a macrophage profile. A high density, radiation-sensitive cell was identified which stimulated the response of PFC precursors in lighter regions of the gradient. The activity of this cell could be replaced using thymus cells. A density profile for the PFC precursor cell was obtained by assaying small numbers of spleen cell fractions in the presence of an excess of the two auxiliary cell types.
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A method for the primary stimulation and measurement of antibody production to sheep red blood cells in vitro has been described. In this system when mouse spleen cells are incubated with sheep erythrocytes large complement-dependent plaques are formed. The number and size of plaques increases from day 1 to day 3 of incubation with an average of 4.4 plaque areas per 1 x 10(6) cells plated at day 3. There is a linear relationship between the number of spleen cells plated and the number of plaques formed. Plaque formation is inhibited by colchicine, actinomycin D, and rabbit anti-mouse globulin. This system offers a possible means for the direct in vitro measurement of the number of cells in a population susceptible to antigenic stimulation by sheep erythrocytes.
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