Mesenchymal stem cells (MSCs) are at the forefront as therapeutic tools for an extensive number of tissue engineering and regenerative medicine applications. MSC differentiation properties have been extensively studied in vitro by this laboratory and many others. The generation and validation of in vivo potency assays would be a valuable tool for the study of cellular properties relevant for in vivo applications. We have developed a unique system, we call the Habitat assay, in which porous ceramic cube carrier loaded with human bone marrow (BM)-MSCs (hMSCs) is subcutaneously implanted into immune-compromised mice. These cells have the capacity to create bone tissue and reconstitute the hematopoietic microenvironment within the "Habitat." These donor-derived hMSCs form bone structures by 3-4 weeks and associate as perivascular MSCs. In this study, we have extensively analyzed data generated with the habitat (ceramic cube in vivo assay) using cells derived from 117 hMSC-donors (iliac aspiration); this analysis provides a validation of the platform as a way to study the in vivo effect of several variables involved in the generation of the bony Habitat. These studies show that passage number and the age of the hMSC donor influence the sequence of in vivo bone formation within the Habitat. These variables have been shown to have an effect on in vitro properties of MSCs; in this study, for the first time, we show these effects to be important on an in vivo setting.
Keywords: bone marrow; extraskeletal bone; mesenchymal stem cells; potency assay.