Although there are many chemicals used for sterilization purposes such as ethanol, hydrogen peroxide, bromine water, mercuric chloride, silver nitrate and antibiotics, sodium hypochlorite solutions have been most widely used. Since NaOCl has a strong oxidizing property which makes it highly reactive with amino acids, nucleic acids, amines, and amides, it is highly effective against all kinds of bacteria, fungi, and viruses. The general reaction between amino acids and NaOCl produces the respective aldehyde, NH4Cl and CO2. Thus, during the sterilization process, direct contact of the tissue with NaOCl depending on it’s concentration, application period and temperature may have a hazardous effect on the health of the tissue. In plant tissue culture studies, high-frequency shoot regeneration which is highly affected by tissue health, is a prerequisite for an efficient gene transformation system and a clonal propagation of plants. The most important treatment prior to culture initiation is the sterilization of the tissue. In plant tissue culture, elimination of microorganisms from the tissue has great importance. From one hand, sterilization process aims to eliminate all microorganisms that can easily grow on the tissue; on the other hand, it should guarantee the tissue’s viability and regeneration capacity. This review has focused on the effects of concentration, application period and temperature of NaOCl using for eliminating microorganisms on the viability and in vitro regeneration capacity of the tissue.