There are many factors creating the internal environment of the building. These factors could be biological, physical, or chemical, and they determine the indoor environmental quality. This contribution investigates the gender differences in occupants' perception on various aspects of indoor environmental quality (IEQ). There are many studies, how the conditions of the indoor environment affect the users' performance, health, or comfort. Generally, women are more likely to have symptoms of SBS, such as fatigue, headache, irritated or dry eyes, nose and throat, and skin symptoms. Assessments of user´s perception were carried out at the Institute of Technology and Business situated in the city of České Budějovice, South Bohemia, Czechia. The panellists (students) indicated their immediate evaluation on eight continuous scales regarding air acceptability, odor intensity, thermal comfort, humidity comfort, visual comfort, color comfort, noise load and total satisfaction. Then the percentage of dissatisfied people (PN) was estimated. The gender differences in perception of indoor environmental quality were analysed. This knowledge is necessary and useful to develop appropriate strategies to create and maintain a sustainable internal environment for education and training. Understanding the perception of the quality of the inner environment by students is essential to increase the performance and productivity of not only students but also the academic staff. The long-term low indoor environment quality can lead to poor productivity, performance and ability to learn. It is also necessary to take into account the health of students and also academic staff in the context of the hygiene of the internal environment.