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

Abstract

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.