High variability in rabbit bone marrow-derived mesenchymal cell preparations

The rabbit has been extensively used for preclinical models, especially in orthopedic applications. One of the more troubling features of this model is the high interindividual variability that is encountered and that requires a careful experimental design with sufficient sample size to make judgments valid. We have processed 241 individual preparations of rabbit bone marrow-derived mesenchymal progenitor cells (MPCs) over the last 3 years and have kept detailed records of the performance of these cells in various assays. This communication details the lack of correlation between the analyzed parameters. Bone marrow was harvested from 4-month-old rabbits; the cells were centrifuged, resuspended, and cultured. When cells reached 80% of confluence, they were removed from the plates with trypsin and assayed for their osteo- and chondrogenic potential. The average yield of the 241 individual MPC preparations exhibited a coefficient of variation of 77. An in vivo implantation assay with porous calcium phosphate ceramic cubes exhibited scores with a coefficient of variation of 65. Lastly, an in vitro assay of alkaline phosphatase enzyme activity exhibited the most variability with a coefficient of variation of 132. All of the cell preparations tested in an in vitro aggregate culture assay underwent chondrogenic differentiation. No relationships between any of these parameters were found. The variability of the results within the different assays is interpreted to be the result of the heterogeneity of the preparations. The lack of correlation between the parameters studied shows the importance of the conditions intrinsic to the different assays. These results serve to emphasize that any experimental design involving rabbit, progenitor cells must include a sufficiently large sample size to allow statistically significant and rigorous conclusions.