Long term growth of factor-producing lymphoid and myeloid cells in serum-free medium.
The ability to grow lymphoid and myeloid cells in serum-free culture medium allows researchers to analyze the factors and mechanisms required for hemopoietic cell growth and differentiation without the interference of undefined serum components. Therefore, we used a serum-free medium, RITC 55-9 that consisted of modified Dulbecco's MEM supplemented with bovine serum albumin (BSA), transferrin (Tf) and insulin (Ins) to culture human T lymphoid (Mo), murine myelomonocytoid (WEHI-3B) and murine interleukin (IL)-3-dependent (32Dcl/H4) cell lines. Mo was maintained in RITC for more than 8 months and had a mean viability of 59% and the same doubling times as in serum-containing medium (SCM). Under these conditions, Mo cells produced hemopoietic colony-stimulating activity that included production of a basophil/eosinophil differentiation factor of similar content to that produced in SCM. WEHI-3B cells grown for more than 12 months in RITC, or for more than 3 months in RITC without Tf and Ins, had a doubling time of 20 h, whereas cells maintained in protein-free RITC showed a 2-fold increase in doubling time then died within 3 months. The IL-3 production by WEHI-3B cells cultured in RITC was higher than the production by cells grown in SCM. When IL-3 was assayed in 32Dcl/H4 cells that had been maintained in RITC for more than 4 months, a lower response to IL-3 was found, an indication that components other than the BSA, Tf and Ins in fetal calf serum are required for optimal cell growth and differentiation.
D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D003470:Culture Media; D006133:Growth Substances / Q000096:biosynthesis; D006801:Humans; D007328:Insulin / Q000494:pharmacology; D007377:Interleukin-3 / Q000502:physiology; D007945:Leukemia, Lymphoid / Q000473:pathology*; D007951:Leukemia, Myeloid / Q000473:pathology*; D008213:Lymphocyte Activation; D051379:Mice; D009930:Organic Chemicals; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D013601:T-Lymphocytes / Q000502:physiology*; D014168:Transferrin / Q000494:pharmacology
D003470:Culture Media; D006133:Growth Substances; D007328:Insulin; D007377:Interleukin-3; D009930:Organic Chemicals; D014168:Transferrin; C042320:colony promoting activity; D012710:Serum Albumin, Bovine