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Colony-forming efficiency is a time-honored metric of the proliferation potential of mesenchymal stem cells (MSCs). This commentary raises a concern about the practice of using colony-forming efficiency as a proxy for cell survival. A recently published study from my laboratory investigated this issue. A marker of cellular aging, CD264, was employed to separate human bone marrow MSCs into populations of CD264 −  cells and culture-matched, aging CD264 +  cells with high and low colony-forming efficiency, respectively. In vitro cell survival was evaluated with a single-cell assay; in vivo survival by bioluminescence imaging of MSCs attached to scaffolds that were implanted ectopically in immunodeficient mice. In our study, in vitro and in vivo survival of the MSC populations was independent of colony-forming efficiency. This finding indicates that caution should be exercised before using colony-forming efficiency as an indirect metric of cell survival. Direct measurement of survival may be required. Awareness of this issue should foster a robust experimental design and, thereby, facilitate the translation of MSC research into clinical practice.

A cautionary tale about the use of colony-forming efficiency as a proxy for the survival of mesenchymal stem cells