A comprehensive analysis of in vitro pumps used in cardiovascular research is provided in this review, with a focus on the characteristics of generated flows and principles of flow generations. The cardiovascular system, vital for nutrient circulation and waste removal, generates complex hemodynamics critical for endothelial cell function. Cardiovascular diseases (CVDs) could be caused by the disturbances in these flows, including aneurysms, atherosclerosis, and heart defects. In vitro systems simulate hemodynamic conditions on cultured cells in the laboratory to study and evaluate these diseases to advance therapies. Pumps used in these systems can be classified into contact and non-contact types. Contact pumps, such as piston and gear pumps, can generate higher flow rates, but they have a higher risk of contamination due to the direct interaction of pump with the fluid. Non-contact pumps, such as peristaltic and lab-on-disk centrifugal pumps, minimize contamination risks, but they are limited to lower flow rates. Advanced pumps including piezoelectric and I-Cor diagonal pumps are focused on improving the accuracy of flow replication and long-term stability. The operational principles, advantages, and some disadvantages of these pump categories are evaluated in this review, while providing insights for optimizing in vitro cardiovascular models and advancing therapeutic strategies against CVDs. The outcomes of the review elaborate the importance of selecting an appropriate pump system, to accurately replicate cardiovascular flow patterns.