The long-term organ culture of tissues from adult Necturus maculosus, the mud puppy.

Abstract

Fragments of Necturus maculosus liver, spleen and kidney were cultured at 25°C in 50% Minimal Essential Medium (MEM) or 50% Leibovitz L-15 Medium (L-15) for up to 49 days. The integrity of tissue structure was evaluated, hepatocyte cell and nuclear volumes were measured, the respiration rates of freshly-isolated and cultured liver fragments were determined, and the mitotic incidences in cultured liver, spleen and kidney were estimated. The addition of adrenalin caused a reduction in the glycogen content of liver cultures, and the subsequent addition of insulin resulted in a net increase in glycogen synthesis. Glycogen levels fell in fragments cultured in L-15, but rose in cultures in MEM. Arginase and ornithine transcarbamylase levels fell gradually throughout a 49-day culture period in L-15. Evidence presented supports the position that the survival of tissues in vitro is related to cell size and respiration rate. These experiments show that N. maculosus is a suitable donor of tissues for long-term organ culture studies on the maintenance and control of tissue-type specific structure and function.