Hybridoma Cell Growth Research Articles

Inhibition of cell division and antibody secretion by murine alpha/beta interferon: effects on plasmacytoma and hybridoma lymphoid cells.

Murine alpha/beta interferon (IFN) inhibited the growth of myeloma cells in vitro. Independently of the cell growth inhibition, IFN also reduced the number of antibody secreting myeloma cells as measured by haemolytic plaque assay. The sensitivity of both MOPC 315 plasmacytoma cells and 4F4 hybridoma cells varied, though not to the same extent, depending on the dose of IFN used and the duration of exposure. Inhibition of PFC activity was observed after one day of IFN treatment while inhibition of cell growth was not detected until Day 2 of incubation. A dose of 20 u/ml IFN had no effect on the growth rate of MOPC 315 cells but with 100 u/ml the inhibition of growth was virtually complete. In contrast, an inhibition of PFC activity was observed at all the concentrations tested. The cell growth and PFC activity of 4F4 hybridoma cells, on the other hand, were both inhibited by IFN when used at concentrations as low as 1.25 u/ml. Incubation with higher concentrations of IFN resulted in a progressive reduction in cell growth and PFC activity of 4F4 cells, however to a lesser degree of inhibition compared to that observed with MOPC 315 cells. For example, although virtually 70% of MOPC 315 PFC could be inhibited by culture for two days in the presence of 100 u/ml, it was necessary to use 1,250 u/ml IFN for 4 days incubation before the similar level of PFC inhibition was achieved with 4F4 cells. IFN treatment resulted in an increase in both cellular volume and protein content and this effect was prevented when IFN was previously neutralised by a specific antiserum. IFN-treated cells also showed an inhibition in the incorporation of 3[H]-thymidine but no alteration in the rate of utilization of 35[S]-methionine, when compared with an equal number of control cells.

In vitro production of high titre monoclonal antibody by hybridoma cells in dialysis culture

Relatively high levels of nutrients and low levels of waste products were maintained in cultures of hybridoma cells by dialysis against growth medium supplemented with fetal calf serum (FCS). This resulted in a greater than ten fold increase in cell density and monoclonal antibody titre compared to conventional batch cultures. Exclusion of FCS from the dialysing growth medium resulted in cell death, indicating that a low molecular weight serum component was necessary for growth. This study indicates that nutrient depletion limits hybridoma cell growth in conventional batch culture and also provides a simple and economical means of producing relatively high titre monoclonal antibody in vitro.

Endothelial cell growth supplement: a cell cloning factor that promotes the growth of monoclonal antibody producing hybridoma cells

The efficiency of endothelial cell growth supplement (ECGS), a commercially marketed extract of bovine neural tissue, human endothelial cell supernatant (HECS) derived from freshly isolated endothelial cells, and feeder layers of murine peritoneal cells (PEC), were compared for their ability to support cell fusion, clonal growth, and monoclonal antibody production of murine hybridoma cells. ECGS at 25–100 μg/ml was similar to a 1:5 dilution of HECS in supporting the growth of hybridoma colonies; both ECGS and HECS were superior to PEC feeder cells. Furthermore, hybridomas cloned in ECGS produced anti-lymphotoxin antibodies. The commercial availability and stability of ECGS together with its ability to support hybridoma growth and monoclonal antibody secretion make ECGS a superior growth supplement for the fusion and growth of hybridoma cells in monoclonal antibody production.

5] Use of human endothelial culture supernatant (HECS) as a growth factor for hybridomas

This chapter discusses the use of human endothelial culture supernatant (HECS) as a growth factor for hybridomas. The growth of hybridoma cells is improved by the presence of feeder cells, particularly when the hybridoma cells are not yet adapted to the culture conditions. This is especially so for cultures at low numbers of cells. The chapter describes that the human endothelial culture supernatant (HECS) contains factor(s) that promote the growth and stability of hybridoma cells and in these respects is superior to the commonly used feeder cells. Human endothelial cells (HEC) were cultured at 37 ° in an atmosphere of 5% CO2 in endothelial cell growth medium (ECGM). The chapter explains the production of HECS and assay of its activity on hybridoma cells. In addition, endothelial cells in serum-flee medium also produced HECS activity. This indicates that the production of the HECS activity did not depend upon interaction of HEC and human serum components. However, serum-free HECS was found to be much more unstable during storage than was HECS produced in the presence of serum.

Growth of hybridoma cells in serum-free medium: ethanolamine is an essential component.

A serum-free medium supplemented with a few growth factors was devised to grow lymphocyte hybridomas. The medium was developed with the hybridoma line MPC11-BL, a fusion product between a mouse plasmacytoma cell line (MPC11TG70na3) and mouse (BALB/c) spleen cells. In the process of developing the medium, ethanolamine was found to be an essential growth factor for the hybridoma. Phosphoethanolamine at 10-fold higher concentration could substitute for ethanolamine. Long-term cultivation of the cells was achieved in the defined medium supplemented with insulin, transferrin, ethanolamine, and selenium. The defined medium supported the growth of various other mouse hybridoma cell lines, mostly at a rate comparable to that observed in a serum-containing medium. After one-step ammonium sulfate precipitation of the spent medium, more than 95% of the protein recovered was immunoglobulin as shown by NaDodSO4/polyacrylamide gel electrophoresis.