大鼠骨髓基质细胞体外定向诱导成骨

将大鼠骨髓单细胞悬液静置培养36h,利用骨髓基质细胞贴壁能力强的特点对其进行纯化和扩增培养。采用爬片培养、HE染色、组化染色以及碱性磷酸酶活性和钙含量测定等手段研究培养骨髓基质细胞的形态、分化和分泌基质情况。结果表明,非诱导培养条件下骨髓基质细胞呈梭形,部分传代细胞中可观察到脂肪细胞和肌细胞。经成骨性诱导培养后,骨髓基质细胞发生明显的形态学变化,碱性磷酸酶活性上升,钙含量增加,最终形成典型的矿化结节。提示培养大鼠骨髓基质细胞具有分化成脂肪细胞和肌细胞的能力,但其分化成骨的潜能最为强大。本实验诱导骨髓基质细胞分化为成骨细胞的模式有可能适用于骨组织工程研究。

Directed differentiation of rat bone marrow stromal cells into osteoblast in vitro

The marrow stromal cells (MSCs) of Wistar rats were isolated from other marrow cells and expanded by applying its strong adherent ability to plastic dish. The morphology, differentiation and secretion of calcified matrix of MSCs were studied by cover slip culture, common histological staining, histochemical staining and the determination of alkaline phosphatase (ALP) activity and calcium content. It was shown that the cultured rat MSCs were spindle-shaped. Adipocytes and myocytes could be found in cultured cells. Under osteogenic induction condition, MSCs changed greatly in appearance, accompanied by the increase of ALP activity and calcium content. After 16 days, mineralized nodular aggregates were observed that indicated the formation of osteoblasts and osteocytes. It can be concluded that the rat MSCs have the capacity to differentiate into osteoblast, adipocyte and myocyte, however, among them, the potentiality of differentiation into osteoblast is the strongest and can be used as seed cells in bone tissue engineering.