Use of an agar culture technique for establishing lymphoid cell lines from Marek's disease lymphomas

Lymphoblastoid cell lines derived from Marek's disease (MD) lymphomas have been established with difficulty by a number of workers. We have compared with conventional liquid culture methods the efficiency of a new technique for establishing lymphoid cell lines in which lymphoma cells were cultured initially in agar medium. Cells from 39/79 lymphomas gave rise to loose lymphoid colonies in the seeded agar after 7 days' incubation at 41 degrees C. Two types of macrophage colony also developed. When lymphoid colonies in agar were transferred to liquid culture, 23/39 gave rise to permanent lymphoid cell lines, compared with 8/33 comparable cultures initiated in liquid medium. Twenty-nine new cell lines have been developed from Rhode Island Red, line 6 and line 7 chickens. All carry T-cell markers and the MD tumour-associated surface antigen (MATSA) and showed variable but low responsiveness to lectin mitogens. The new cell lines, when first established, consisted mainly of small, lymphocytoid cells, but, after varying times, these changed into typical lymphoblastoid lines and an increased expression of an embryonic antigen was associated with this change. The lymphocytoid line cells were more slowly growing and density-dependent than were the lymphoblastoid cells, and lymphocytoid lines grew better at 41 degrees C and lymphoblastoid lines better at 37 degrees C. MD virus could be rescued from some of the lines but others appeared to be true non-producers.