摘要
目的研究犬骨髓基质干细胞 (BMSCs)体外成骨活性和在犬体内异位成骨能力 ,探讨其作为骨组织工程种子细胞的可能性。 方法抽取成年犬骨髓 ,梯度离心法获取单个核细胞 ,条件培养液体外培养、诱导、扩增后 ,进行组织化学、免疫细胞化学染色 ,透射电镜下观察其成骨活性。将培养的第 3代细胞与藻酸钙所形成的复合物植于犬皮下 ,3月后行组织学检测。 结果体外培养的BMSCsVonKossa染色可见钙结节 ,AKP染色及I型胶原、Osf2 /Cbfα1、骨钙素免疫细胞化学染色先后呈现阳性 ;透射电镜下见基质钙化 ;组织学检测提示细胞 -藻酸钙复合物体内培养 3月后形成骨样组织。 结论BMSCs在大量扩增的同时 ,在一定条件下能向成骨细胞分化 ,细胞与藻酸钙复合物能在具免疫力的哺乳动物皮下形成骨样组织。
Objective To study the osteogenesis ability of canine bone marrow stem cells (BM SCs ) cultured in vitro or in vivo and the probability that they may be used as seed cells for bone tissue engineering. Methods The BMSCs were separated by gradient centrifugation on Percoll,culture-expanded and induced with the conditional medium.The phenotype of the BMSCs was evaluated by transmission electron microscopy and histochemistry or immunocytochemistry.After BMSCs culture and expansion,the induced passage 3 cells and calcium alginate were mixed to form a complex.Then,the complex was auto-implanted into the subcutaneous part of the dog to reconstruct new tissues. Results Histochemical or immunochemistry studies showed positive stain of Von Kossa,alkaline phosphatase(AKP),Core-binding factor alpha subunit 1(Osf2/Cbfα1)?type Ⅰ collagen and osteocalcin(OCN).The extracellular matrix calcification was observed under transmission electron microscopy.After 12 weeks of the complex implanted,bone-like tissue were formed. Conclusion BMSCs have the osteoblast-like structure and function when cultured in vitro and in vivo .They might be the ideal seed cells for bone tissue engineering.
出处
《上海第二医科大学学报》
CSCD
2003年第4期289-293,共5页
Acta Universitatis Medicinalis Secondae Shanghai
基金
国家 973资助项目 (G1 9990 5430 0 )
