Organizational culture is an important construct that affects both individual and organizational related process and outcomes. The study aimed to assess organizational culture as perceived by nursing personal at Benha and Shebin Elkom University Hospitals. A descriptive comparative design was utilized to meet the aim of this study .The sample of this study were included 400 stuff nurse (200 of them were working at Benha University Hospital and other 200 were working at Shebin Elkom University Hospital) head nurses were 60 (30 of them were working at Benha Hospitals and other 30 were working Shebin Elkom Hospitals. Tools for data collection One tool were used for data collection of this study, organizational culture assessment questionnaire (OCAQ),  it will developed by the researcher after reviewing  the  related  literature to assess organizational culture It will consist of  two parts: Part (1): demographic data as (age, marital status, education, etc). Part (2): Include 41 questions to assess organizational culture the major items they are namely: dominant characteristics, organizational leadership, management of employees, organization glue, strategic emphases, criteria of success and Relationship of the patient to organization. The result of the present study revealed that staff nurses perception at Benha hospital was higher than staff nurses at Shebin Elkom Hospital related organization culture, there was a highly statistical significant difference between nurses at Benha Hospital & Shebin Elkom Hospital regarding their perception about (dominant characteristics, organizational leadership, management of employee, organization glue, strategic emphases, criteria of success and relationship of the patients to organization), There was statistical significant difference between head nurses at Benha Hospital & Shebin Elkom Hospital regarding their perception about dominant characteristics, There was no statistical significant difference between head nurses at Benha Hospital & Shebin Elkom Hospital regarding their perception about (organizational leadership, management of employee, organization glue, strategic emphases, criteria of success and relationship of the patients to organization).The study recommended continue motivation for nurses through development of human teamwork, employee commitment, and concern for employee, In-service training and education programs must be a continuous process for increasing cognitive abilities of staff underlying decision- making, and culture competencies.