Poultry semen has high spermatozoa concentration and needs to be extended (diluted) for efficient artificial insemination; extenders containing egg yolk often have various limitations to their use. This study assessed chicken egg yolk plasma (EYP) as replacement for chicken whole egg yolk (EY) in semen extenders. The preservative potential under room and cold storage and its influence on fertility and hatchability in breeder chicken flock was also assessed. Ten broiler breeder cocks and one hundred hens were used for the study. Semen ejaculates from the cocks were pooled and divided into five portions. One portion each was extended with egg yolk plasma (EYP); phosphate buffer saline (PBS); egg yolk (EY) +PBS or EYP +PBS while the fifth unextended portion termed fresh undiluted semen (FUS) served as the control. The ratio of EYP and EY to PBS was 1:4. The semen samples were evaluated at 1, 2 and 3 hours after extension under room temperature and at 24, 48 and 72 hours after cold storage (4℃). The hens were inseminated with the freshly extended semen or with cold preserved semen, each for three weeks. Fertility and hatchability of the eggs were recorded on 18th and 21st day, respectively post setting. At room and cold storage temperatures, the quality parameters of the semen significantly reduced as the holding time increased. The EYP groups had the highest spermatozoa motility of 55.33% at 72 hours of cold storage. Freshly extended semen with EYP+ PBS had higher quality and fertilizing potentials which resulted in increased egg fertility (87.49%) and hatchability (84.95%). Cold preserved EYP + PBS semen resulted in significantly higher fertility (75.66%) and hatchability (90.90%). It was concluded that egg yolk plasma could conveniently replace whole egg yolk in chicken semen extender resulting in improved sperm viability and egg hatchability.