Development ofachemically defined serum- andprotein-free mediumforgrowth ofhumanperipheral lymphocytes

A chemically defined, protein-free medium (designated CFBI1000, whereCFBI= Clayton Foundation Biochemical Institute) thatsupports humanperipheral lym- phocyte proliferation hasbeendeveloped. This medium allows exploration ofindividual metabolic differences byvarying the mediumcomposition aswell asproviding abasetoexplore further themechanisms oflymphocyte activation inasystem initially freeofaddedmacromolecular species other than mitogen. Theperipheral blood lymphocyte isanideal system formetabolic studies because itiseasily obtained, isaprimary resting cell that canbeactivated toproliferate, andpresumably reflects boththegenetic makeup andbiochemical environmen- talhistory oftheindividual atthetimethecells wereformed. Examination oftheroleofvarious factors inlymphocyte activation andsubsequent events maybesimplified bythe utilization ofamediumthatisprotein-free andchemically dermed. TheCFBI1000 mediumsupports thegrowth response ofhumanperipheral lymphocytes tomitogen asmeasured by (3H)thymidine incorporation toanextent comparable toother mediausedwidely inassessment oflymphocyte proliferation. Theeffects ofalterations inmediumupongrowth ofcells taken directly fromanindividual subject havebeeninvesti- gated foranumber ofyears inthis laboratory asanapproach totheindividualized assessment ofmetabolic andnutritional status. Theready availability oflymphocytes fromhuman peripheral bloodrelative toother cells andtheir known activation andblastogenesis inculture media containing fetal calf serumledtotheir selection aspotential cells for this work (1,2).Theprimary resting state oftheTlymphocyte canbe interrupted bythepresence ofantigen inassociation with major histocompatability complex or,morecommonly incell