In vitro cultivation of ovaries and ovarian tissue from the red imported fire ant, Solenopsis invicta Buren

A method for in vitro cultivation of ovaries and ovarian cells from Solenopsis invicta has been developed. Whole ovaries were kept alive, in Schneider's Dropsophila medium (pH 6.7) and Hink’s THH-FH medium (pH 6.1, 7.0), for up to six weeks without refeeding. Cells from physically and chemically dissociated ovaries were kept alive for up to 15 days. Very little evidence of mitosis was observed in cell cultures. TNM-FH, pH 7.0, supported cell cultures which were superior to those obtained in the other tested media. The addition of fetal bovine serum, bovine serum albumin, chick embryo extract, whole egg ultra filtrate, and glycylhistidyl-lysine to this medium did not induce growth or longevity greater than that observed in control media, which contained only fetal bovine serum. Overnight refrigeration of whole ovaries in 0.5 mg/ml collagenase solution, followed by 60 minutes incubation at 28o C and physical dissociation, gave a cell yield which was superior to that obtained by other methods of tissue disruption.