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

Abstract

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.

Bone Marrow Concentrate Mesenchymal Stromal Cells Do not Correlate With Nucleated Cell Count or Colony Forming Units

“…5,31,32 This suggests that the CFU assay is a qualitative test of replicative health of a cell as originally described 33 and not a method to quantify MSCs in BMC. We agree with O'Connor 34 that colony-forming efficiency should not be used as an indirect metric of cell survival, and the continued misuse of the CFU assay has the potential to slow the translation of MSCs research into clinical practice.…”

Characterization of a Stemness-Optimized Purification Method for Human Dental-Pulp Stem Cells: An Approach to Standardization

“…The colony forming capacity is an established proxy for the proliferative potential of SCs [ 138 ]. Colony-forming capacity was found to be highest in (+) cells, followed by (c) cells.…”

Antitumor effects of new glycoconjugated PtII agents dual-targeting GLUT1 and Pgp proteins

“…Colony formation is one of the common methods to determine clonogenic survival. 44,45 Generally, formed colonies that consist of at least 50 cells being counted are defined as the ability of the cells to maintain their clonogenic capacity. As shown in Fig.…”