Background: Accurate blood typing is essential for blood transfusions, and requires the constant evaluation and maintenance of ABO and D blood grouping reagents. In the present study, we developed cryopreserved red blood cell (RBC) panels and evaluated their feasibility as a standard reference material to verify the quality of ABO and D blood grouping reagents in Korea. Methods: RBC units obtained from healthy donors were cryopreserved using a high-glycerol method. A total of 400 sets of RBC panels were prepared, composed of blood group A (N=5), B (N=5), O (N=10), AB (N=4), Rh D-positive (N=4), Rh D-negative (N=5), and weak-D (N=1), and 200 sets of RBC subgroup panels composed of A2, A2B, A2B3, A1B3, and B3, and A2, A2B, A2B3, A1B3, and A3B (N=1, each). Quality assessment of the cryopreserved RBC panels before and after cryopreservation was performed by measuring their sensitivity, specificity, avidity, and potency titers. Results: Our cryopreserved ABO and D RBC panels had a sensitivity and specificity of 100% to existing monoclonal blood grouping reagents, regardless of blood type and cryopreservation time. There were no significant differences in the avidity time and potency titers of the cryopreserved RBCs before and after 6 or 12 months of cryopreservation. Conclusions: The quality parameters measured here suggest that our newly developed cryopreserved RBC panels were reliable for use as a standard reference material for the performance evaluation of anti-A, -B, and -D blood grouping reagents.