Improved Efficiency of Hybridoma Ascites Production by Intrasplenic Inoculation in Mice

Intrasplenic inoculation of small numbers (104) of antibody-producing hybridoma cells into unprimed BALB/c mice resulted in high-titered ascites. By intrasplenic injection, the low number of cells needed to produce ascites was 2–3 logs fewer than that required by ip inoculation. Hybridoma cells suspended in the ascitic fluid could be reestablished in vitro. Furthermore, cells obtained directly from individual, cloned colonies or from previously frozen samples also resulted in high-titered ascites, if the cells were inoculated intrasplenically. This method reduces time and expense required to grow large numbers of hybridoma cells for ip-derived ascites production and allows ready recovery of frequently unstable frozen-thawed cells.