Tumour Promoters/Protein‐Kinase C Activators Augment the Survival and Function of Human Monocyte‐Derived Macrophages in Long‐Term Cultures

Abstract

In this study the effect of various Protein kinase C (PKC) activators/tumour promoters on the maturation and activity of human peripheral blood monocytes was examined. Monocytes were cultured in the absence or presence of various PKC activators for up to 2 weeks, and examined for the number of adherent cells, expression of myeloperoxidase enzymes, CD14 antigens, mannose/N-acetylglucosamine (Man/GlcNAc) receptors, and the production of TNF-alpha. The presence of PKC activators in cultures of monocyte-derived macrophages (HuMoDM) prevented the loss in the number of initially plated monocytes, otherwise observed in long-term tissue cultures with time of incubation. This effect of PKC activators on monocyte survival was diminished in the presence of PKC inhibitors. HuMoDM obtained in the presence of PKC activators maintained a normal differentiation pattern, as was evident by the loss of granular myeloperoxidase enzymes and CD14 antigens, and the acquisition of membrane Man/GlcNAc receptors. HuMoDM which differentiated in the presence of PKC activators also released TNF-alpha in comparable amounts to freshly harvested human monocytes. PKC activators/tumour promoters augmented the viability of long-term cultures of human monocyte-derived macrophages. Such macrophages may facilitate cell and molecular biology studies of differentiated human macrophages.

Regulation and specificity of MNDA expression in monocytes, macrophages, and leukemia/B lymophoma cell lines

“…In addition, TPA at the nanomolar concentration promotes the viability and recovery of long-term cultures of human monocyte derived macrophages [Markovich et al, 19941. This treatment of adherent monocytes also facilitates the ability of the cells to secrete TNFa (Markovich et al, 1994). The effectiveness of TPA to eliminate the MNDA mRNA signal in monocytes was observed following treatment at the micromolar or nanomolar levels (Fig.…”

The LINC-less granulocyte nucleus

“…In other experiments, blood monocytes were enriched from the mononuclear fraction by adherence to glass chamber tissue culture slides (Keisari, 2005), washing away lymphocytes and other non-adherent blood cells. Some of the monocyte chamber slides were made 2 nM TPA in RPMI 1640 plus 10% heated fetal calf serum and incubated for ~2 weeks to induce macrophage differentiation (Keisari, 2005; Markovich et al, 1994). Examples of confocal immunostaining results of the reconstituted “buffy coat” are presented in Figure 7.…”