With regard to the bone-regenerative capacity, bone marrow stromal cells (BMSC) can still be termed the “gold standard.” Nevertheless, neonatal stromal cells from cord blood (CB) feature advantages concerning availability, immaturity, and proliferation potential. The detailed gene expression analysis and overexpression of genes expressed differentially provide insight into the inherent capacity of stromal cells. Microarray and qRT-PCR analyses revealed closely related gene expression patterns of two stromal cell populations derived from CB. In contrast to the CB-derived cell types, BMSC displayed high expression levels of BSP, OSX, BMP4, OC, and PITX2. Lentiviral overexpression of BSP but not of OSX in CB-cells increased the capacity to form a mineralized matrix. BMP4 induced the secretion of proteoglycans during chondrogenic pellet culture and extended the osteogenic but reduced the adipogenic differentiation potential. BMSC revealed the typical osteogenic gene expression signature. In contrast, the CB-derived cell types exhibited a more immature gene expression profile and no predisposition towards skeletal development. The absence of BSP and BMP4—which were defined as potential key players affecting the differentiation potential—in neonatal stromal cells should be taken into consideration when choosing a cell source for tissue regeneration approaches. 1. Introduction With respect to the regeneration of cartilage or bone after tumor resection, accidents, or due to diseases affecting the skeleton, there is, great need for tissue-engineered bone. The cellular component of these approaches has been in the focus of interest for many years. The first described [1] and therefore the best studied nonhematopoietic stromal cell type derives from bone marrow (BM). The in vivo bone forming potential—including recruitment of hematopoietic cells of recipient origin—of these bone marrow stromal cells (BMSC) after transplantation on a hydroxyapatite scaffold was reported by several groups [2, 3]. The potential risks associated with the bone marrow donation made other sources of stromal cells, for example, adipose tissue or peripheral blood, attractive alternatives. Due to its immaturity compared to adult bone marrow, neonatal cord blood (CB), which can be collected noninvasively and without ethical concerns, can be regarded as a proper source of neonatal stromal cells with potential clinical relevance in the future. Cord blood contains at least two distinct populations of nonhematopoietic stromal cells with comparable proliferative potential [4], which were termed
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