犬骨髓基质干细胞诱导为成骨细胞的实验研究

目的：研究骨髓基质干细胞（Bone marrow stromal cells,BMSCs)在体外培养，诱导后的成骨活性，探讨BMSCs作为骨组织工程种子细胞的可能性和可行性。方法：抽取成年犬骨髓，用梯度离心法获取单核细胞，经条件培养液体外诱导培养后，进行组织化学、免疫组化、原位杂交检测，并在透射电镜下观察其成骨活性。结果：第1代细胞Alkaline phosphatase(AKP)染色、Core-binding factor alpha subunit 1(Osf2/Cbfal)、Osteopotin(OPN),I型胶原的免疫细胞化学检测和I型胶原的原位杂交结果开始呈现阳性，第3代细胞Bone Sailoplain(BSP),Osteocalcin(OCN)免疫细胞化学染色开始呈现阳性，透射电镜显示细胞外有胶原分泌和钙盐沉积。结论：BMSCs不仅能大量扩增，且在一定条件下可向成骨细胞分化，是骨组织工程理想的种子细胞。

Experimental Study of the Inducing BMSCs into Osteogenesis

OBJECTIVE: To study the bone tissue osteogenesis ability of bone marrow stromal cells (BMSCs) in vitro and in vivo. METHODS: The BMSCs were separated by gradient centrifugation on Percoll, culture-expanded and induced with a conditional medium. The cells were examined by transmission electron microscopy and histochemistry or immunochemistry stains technique including AKP,Cbfa1,OCN, OPN, BSP and collagen type I. The mRNA of collagen type I in cultured cell was also examined by in situ hybridization. RESULTS: Histochemical studies showed positive stain of AKP,Cbfa1, OCN, OPN,BSP and collagen type I,and in situ hybridization also showed the positive express of mRNA of collagen type I. The excretion of collagen and deposition of calcium salt were observed under transmission electron microscopy. CONCLUSION: BMSCs cultured in conditional medium shows osteoblast-like functions in vitro, which provides an ideal seed cells for bone tissue engineering.