Tthe desirable cultivars of date palm (Zaghloul and Samany) beginning from segments of fruits flesh to obtain initial callus, compact callus and meristematic center cells. For this purpose, a study was conducted to define micropropagation protocol based on the cell totipotency to obtains formation of callus. Sodium hypochlorite was very effective for surface sterilization of explants. The best method of surface sterilization was achieved by using treatment containing NaOCl 70% that produced the lowest percentage of contamination 1.33% for Zaghloul and 1.58% for Samany explants by time expanding during sterilization period from 10 to 30 mint. It was observed that the highest percentage of callus was produced at Khalal stage than other stages of fruit development of Zaghloul and Samany cv., when culturing fruit flesh in MS medium containing 100mg/1 of 2,4-D or 30mg/l of Picloram (Pic). The effect of physiological stage of fruit and growth regulators on callus browning was investigated. The results indicates that the treatment supplemented with Pic (30mg/l) produced the highest amount of total soluble phenols in callus 3.73 mg/100g F.W for Zaghloul at Rutab stage and 3.42 mg/100g F.W for Samany at kimri stage. On the other hand, there were significant differences between all stages on total soluble phenols of callus in Zaghloul. While there was no any significant difference among Khalal and Rutab stage on callus formation from Samany explants. The stages of initial callus formation and callus percentages of date palm (Phoenix dactylifera L.) were studied from the 2-10 months-old cultures in MS medium containing 2,4-D or Pic at different concentrations. The study revealed that the compact callus was formed from the friable callus and meristematic center cells. The number of these center cells were unequally distributed inside the compact callus.