Development of a serum‐free medium which supports the long‐term growth of human and nonhuman primate lymphoid cells

Abstract The ability to grow lymphoid cells in serum‐free media affords the advantage of separately analyzing those components found to be involved in proliferation and differentiation. Iscove's medium (IMDM) supplemented with bovine serum albumin or casein, cholesterol, ferrous chloride, insulin, β‐mercaptoethanol, L‐α‐phosphatidylcholine, and transferrin supported the long‐term proliferation of a gibbon ape lymphoma T‐cell line, MLA144. These cells continue to produce Interleukin 2 (IL‐2, T‐cell growth factor) constitutively in the serum‐free medium. IL‐2‐dependent human T cells initiated and maintained in culture in serum‐free medium containing IL‐2 have continued to replicate for over 3 months with two population doublings every 3 to 4 days. A normal, IL‐2‐dependent marmoset T‐cell line, OH‐1, also proliferated on the serum‐free medium when supplemented with IL‐2. Several established primate B‐cell lines which do not require IL‐2 for growth were able to proliferate in the serum‐free medium. These B‐cell lines included B95‐8, an Epstein‐Barr virus (EBV)‐transformed marmoset cell line, HuCo/R‐H, a human cord B‐lymphocyte line transformed with EBV, and Namalwa, an EBV‐positive B‐cell line established from a Burkitt's lymphoma. B95‐8 cells grown on serum‐free medium showed high levels of EBV antigen‐positive cells after induction with 12‐O‐tetradecanoyl‐phorbol‐13‐acetate (TPA).