Multipotent mesenchymal stromal cells also known as mesenchymal stem cells according to the signal from the damaged tissue have the ability to differentiate into several types of specialized cells forming tissues of mesodermal origin such as bone, cartilage, tendon, skeletal muscle or adipose tissue. This ability of MSC is used in regenerative medicine. For the clinical and experimental purposes in order to increase the amount of MSC their ability to proliferate in vitro is used. The best-known source of mesenchymal stem cells is the adipose tissue and bone marrow which is the most common source material used for primary culture. This paper presents the next steps of culturing mesenchymal stem cells in vitro.