An altered cellular glucocorticoid (GC) sensitivity is associated with several pathophysiological conditions such as asthma, diabetes, or rheumatoid arthritis. Several bioassays have been developed and employed to assess cellular GC sensitivity of peripheral blood mononuclear cells (PBMC), but correlations between these have rarely been investigated. We have compared four mitogen-based assays and an FK506 binding protein 51 (FKBP51) mRNA induction assay, using ten controls and a GC-resistant patient. The mitogen-based assays were performed using either diluted whole blood or isolated PBMC, and showed relatively large assay variations for the parameters maximal effect and half-maximal effect concentration. The FKBP51 assay showed smaller intra-assay and within-individual variation compared with the mitogen-based assays. The whole blood-based mitogen assays and the FKBP51 assay clearly discriminated the GC-resistant patient from the controls but, in contrast to expectations, both PBMC-based mitogen assays did not. The GC-induced FKBP51 mRNA increase in PBMC may be an alternative to determine an altered individual GC sensitivity with several advantages as compared with mitogen-based assays, such as the use of unstimulated PBMC, and a better intra- and inter-individual reproducibility.