Characterization of biological response modifier release by human monocytes cultured in suspension in serum-free medium.
Human monocytes have been shown to be critical immunoregulatory cells for a variety of frequently measured in vitro human immune functions and are secretors of potent biologic response modifiers ( BRMs ). These BRMs , such as interferon (IFN), colony stimulating factor (CSF) and prostaglandin E (PGE), may play important roles in the host immune response to cancer. A new approach for culturing circulating peripheral blood monocytes has been developed to retain the native characteristics of these cells, avoiding possible alteration or activation by adherence. The ability of elutriator-purified human monocytes to secrete IFN and PGE was examined under conditions of suspension culture as well as after adherence, and no difference in secretion of these BRMs was noted. In contrast, Teflon-cultured monocytes demonstrated a significantly enhanced CSF release over culturing in polystyrene plates. A new serum-free medium has also been developed, and monocytes cultured in vitro in this medium showed a 5-fold increase in IFN release, and up to a 72% increase in CSF release when compared to optimal standard culture medium (containing 10% AB serum) and an increased stimulation index for PGE release. By these procedures, hundreds of millions of highly purified human monocytes can be sterilely isolated in suspension and cultured in suspension in serum-free medium with retention of BRM-releasing capabilities. This system should permit more detailed molecular studies of monocytes (in which serum can be an impediment) and may also facilitate clinical therapy studies involving the in vivo transfer of monocytes activated in suspension in vitro.
D001790:Blood Physiological Phenomena; D002448:Cell Adhesion; D002452:Cell Count; D002470:Cell Survival; D002478:Cells, Cultured; D003115:Colony-Stimulating Factors / Q000096:biosynthesis; D003470:Culture Media; D006801:Humans; D007158:Immunologic Techniques / Q000295:instrumentation; D007372:Interferons / Q000096:biosynthesis; D009000:Monocytes / Q000276:immunology / Q000378:metabolism* / Q000502:physiology; D015846:Monokines; D011458:Prostaglandins E / Q000096:biosynthesis; D014176:Protein Biosynthesis
D003115:Colony-Stimulating Factors; D003470:Culture Media; D015846:Monokines; D011458:Prostaglandins E; D007372:Interferons