Objectives: To isolate probiotic bacteria from human breast milk and test their   health benefits as cholesterol reduction and  human DNA protection. Methods: Potential probiotic bacteria were isolated from human milk.  The probiotic properties were tested namely their ability to survive acidic pH, tolerate bile salts and  antimicrobial activities. Serum cholesterol reduction ability and cytoprotective effect on Hep-2 cells were studied. Results: A total of 48 different isolates were purified from 12 breast milk samples. They all tolerated   bile salts while 81% were proved to be resistant to acidic pH and had  antimicrobial effects against different pathogens .  Three isolates showed high ability to reduce cholesterol from 19.4-22.6%. This percentage of cholesterol reduction was improved in presence of bile salt to be 48.7-54.7%. Four isolates were proved  for  a protective effect against the H₂O₂ cytotoxicity by decreasing tail moment up to 2.8 and 15.6% DNA concentration in the tail of the examined cells. Phenotypic characteristics and 16S rDNA sequence analyses were used to identify the promising isolates as Pediococcus pentosaceus, Enterococcus durans, and Enterococcus faecium strains. Conclusion:  Probiotic  strains isolated from human milk in the study had a dual beneficial role; human health benefit as well as cytoprotective activities.