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Multiple studies have revealed that mesenchymal stem cells (MSCs) isolated from adult bone marrow are the precursors of adipocytes and osteoblasts and will differentiate into osteoblasts in vitro. Unfortunately, MSCs isolated from bone marrow are heterogeneous and their functional status (e.g. self-renewal, differentiation, and dormancy) is difficult to evaluate, and the data are often inconsistent. Thus, much controversy remains regarding the mechanism(s) of how MSCs are mobilized to produce mature osteoblasts and their niche within bone, and whether there is a hierarchical relationship between mature osteoblasts and adult MSCs. This chapter provides an overview of the tissue sources for MSCs, methods of MSC isolation, and differentiation of MSCs into cells of the osteoblastic lineage. It further reviews a variety of histologic methods that have been used to analyze murine and human MSCs in vivo or in tissue culture. We review the use of critical point drying, freezing, and cryostatic sections for analysis of ultrastructural features and autoradiography to quantify MSCs. Representative techniques for in situ hybridization, immunohistochemistry, transmission electron microscopy, in situ terminal uridine deoxynucleotidyl transferase (TUNEL) staining, and in situ detection of mineral (von Kossa, Alizarin red) are also discussed.Q:
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