The action of cytostatic agents on the chromosomes of human leukocytes in vitro (preliminary communication).

Four humndred aml(l ten c’Imitumr(’s of normllal Peril)heral human i(’umkocytcs vt’re prel).ure(i alit1 treate(l accordimig to the mnt’tho(l of Nlooriieati et al. mb vitI1 slight miiodihcatioiis. Time 1)100(1 was taken fromu 46 1)atidnts sumifering from nonkumkemnic internal (liseases, none of these having ireviously receivv(l cytostatic agents or irradiation. 0.02 ml. hactophytoheniagglumtinin \1 ( 1)ifco ) were (t(l(Id(l to the hep:trinized 1)100(1 sammiples, uvhich vere then kept for al)oumt 30 nminumtcs tt a temuperattmrt’ of 2-4 C. Thc cumituire mmmediumm ( total ammu)umnt 10 iiiI. ) was 30) P ’ cent aumtologous plasilma. 70 i) r cent TC 199 ( i)ifco ) Penicillin. 0.26 mmmg./miml., and dihvdrostreptonivcin, 0.2 mmmg./nml., were added to suppress l)adterial (.‘Ontamlliriants. The cultures originally containing 1 O) leimkocytcs /mni . niediumu were induml)ate(l for 72 hours at a temmmperatuire of 37.0 ± 0.2 C. Twenty-fouir houmrs i)efore fixatiomi, one of the water soluble cvto: tatic agents to be investigated was ,idded to the cultumres. Control cumltumres, which received only the solvent, were imwluded in each series of experimnents. At 4 houmrs before fixation. 0.004 mug. (lesacetylmllethvkoichicin (colcemide) was added for each miii. of mediumimm. H,vpotonic treatment with distilled water 1:3 was carried omit for exactly 10 mmminuites. followed l)y 5 minutes of centrifugation. Time miietaphases were