Abstract

Bone marrow stromal cells (BMSCs) can differentiate into osteocytes and form mineralized matrices in culture conditions. This chapter describes the structural characteristics of in vitro-cultured bone tissue, which includes mineralized matrices and differentiated cells, formed by rat BMSCs. In addition, I introduce a three-dimensional (3D) visualization method for cultured bone tissue using confocal laser scanning microscopy. This 3D visualization showed that the in vitro-constructed tissue contains layers of mineralized matrices with round cells in the matrix lacunae and osteoblastic cells on the mineralized matrices. Type I collagen fibers were well integrated within the mineralized matrices. Mineralized matrices near the culture dish surface did not contain deposited collagen fibers; however, in the upper part of the matrices, calcium was deposited as small particles along with collagen fibers. The structure of the in vitro-mineralized tissue formed by BMSCs is comparable to that of normal bone tissue formed in vivo. This 3D visualization method will aid in the in vitro construction of 3D bone tissue for tissue engineering applications.

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