Research progress on the impact of pre-analytical variables on RNA quality in biobank tissue samples

<?A3B2 tlsb=-.01w?>Biobanks represent irreplaceable core resources for translational medicine and precision medicine research. The quality of preserved tissue samples directly determines the reliability and reproducibility of downstream molecular analyses, among which RNA integrity is widely acknowledged as a pivotal quality indicator of tissue specimens. However, RNA quality is significantly influenced by numerous pre-analytical variables throughout the entire lifecycle from sample collection to analysis. Elucidating the mechanisms underlying these variables provides essential theoretical basis and practical guidance for the development of standardized biobank protocols. This paper systematically reviews key pre-analytical variables that influence RNA quality in tissue samples. These variables are categorized into two phases: pre-acquisition and post-acquisition phases. The pre-acquisition phase mainly encompasses donor characteristics (age, BMI, disease status, and medication history), tissue factors (organ origin, histological features, tumor grade, tumor location, and clinical stage), and surgical variables (surgical approach and warm ischemia time). The post-acquisition phase focuses on cold ischemia time, preservation temperature, storage duration, freeze-thaw cycles and the selection of RNA stabilizers. The review elaborates on the effects of these factors on RNA integrity across different tissue types, and summarizes the controversial findings and underlying molecular mechanisms in current studies. Based on comprehensive analyses, this review proposes that biobanks should implement dynamically optimized and full-lifecycle quality control strategies tailored to specific tissue types and research objectives. It is recommended to strictly regulate the pre-analytical window periods, establish a full-chain temperature monitoring system, adopt aliquot storage to minimize freeze-thaw damage, and apply the Standard PREanalytical Code (SPREC) for standardized recording and traceability of key pre-analytical variables. Furthermore, the combined application of multiple RNA quality evaluation systems (such as RIN, RNA IQ, and RQI) and regular sampling quality control is recommended to enhance the accuracy of RNA quality assessment. This review provides a robust theoretical foundation for the development of standardized, high-quality biobanks and facilitates the efficient utilization of biospecimens in clinical translation and scientific research. <alternatives id="alt3"> </alternatives>