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Newly ovulated eggs from immature rats treated with PMSG and HCG were `inseminated' in vitro with spermatozoa recovered from the cauda epididymidis of mature males. A modified Krebs-Ringer bicarbonate solution, supplemented with glucose, sodium pyruvate, sodium lactate, bovine serum albumin and antibiotics, was used. Eggs and spermatozoa were incubated in 0\m=.\4ml medium under mineral oil at 37\s=deg\C in a gas phase of 5% CO2 in air. Sperm penetration through the zona pellucida started 5 to 5\m=1/2\hr after insemination and the transformation of sperm head(s) into the male pronucleus occurred about 2 hr later. Most of the penetrated eggs were at the two-cell stage 30 to 35 hr after insemination. The percentage of eggs undergoing fertilization was 88\m=.\7%(316/356) 7 to 20 hr after insemination and that of fertilized eggs at 30 to 46 hr after insemination

FERTILIZATION OF RAT EGGS IN VITRO BY EPIDIDYMAL SPERMATOZOA AND THE DEVELOPMENT OF EGGS FOLLOWING TRANSFER