This review article explores the pharmacokinetic characteristics of mesenchymal stem/stromal cells (MSCs) and their challenges in translational applications. MSC therapy has shown promise in treating various critical conditions, but it faces limitations in detecting the dynamic biodistribution, persistence, and ultimate fate of MSCs in patients. The relationship between MSC pharmacokinetics and therapeutic efficacy is crucial for patient stratification and precise treatment regimens. Advanced imaging and tracking technologies are essential to address these challenges. The review discusses the kinetic properties, key regulatory molecules, different fates, and detection methods of MSCs, emphasizing the importance of integrating pharmacokinetics and efficacy. Understanding these challenges can improve the clinical efficacy of MSC therapies and accelerate their commercialization. The article also covers the landscape of clinical MSC therapies, the molecules governing MSC motility during homing, and the target organs of MSCs after systemic administration. Additionally, it examines the fate of MSCs post-transplantation, including apoptosis, autophagy, differentiation, ferroptosis, phagocytosis, and senescence.This review article explores the pharmacokinetic characteristics of mesenchymal stem/stromal cells (MSCs) and their challenges in translational applications. MSC therapy has shown promise in treating various critical conditions, but it faces limitations in detecting the dynamic biodistribution, persistence, and ultimate fate of MSCs in patients. The relationship between MSC pharmacokinetics and therapeutic efficacy is crucial for patient stratification and precise treatment regimens. Advanced imaging and tracking technologies are essential to address these challenges. The review discusses the kinetic properties, key regulatory molecules, different fates, and detection methods of MSCs, emphasizing the importance of integrating pharmacokinetics and efficacy. Understanding these challenges can improve the clinical efficacy of MSC therapies and accelerate their commercialization. The article also covers the landscape of clinical MSC therapies, the molecules governing MSC motility during homing, and the target organs of MSCs after systemic administration. Additionally, it examines the fate of MSCs post-transplantation, including apoptosis, autophagy, differentiation, ferroptosis, phagocytosis, and senescence.