【摘要】 为了研究犬骨髓间充质干细胞(can ine m esenchym a l stem ce lls,cM SC s)的生长特点和在诱导条件下的成骨特性。使用密度梯度法分离成年犬骨髓间充质干细胞进行培养,保留贴壁细胞传代,观察,以地塞米松、β甘油磷酸钠、抗坏血酸为成骨诱导剂。利用倒置光学显微镜和透射电镜观察细胞形态特征,四甲基偶氮盐(M TT)比色测定增殖,用碱性磷酸酶(A lka line phosphatase,ALP)活性及骨钙素(O steoca lc in,OCN)含量来研究细胞分化情况。形态学观察表明,cM SC s贴壁细胞呈集落生长,有成纤维细胞样外观,透射电镜可见成骨诱导后cM SC s具有分泌型细胞的特征;推测成骨诱导剂可促进骨髓间充质干细胞成骨,表现为ALP活性、OCN含量明显升高。本实验表明所培养的cM SC s保持了未分化状态,并在成骨诱导剂的作用下可向成骨细胞分化。更多还原

【Abstract】 To investigate the growth and osteogenesis characteristics of cultured canine mesenchymal stem cells(cMSCs)under osteogenic induction.We found the cMSCs were isolated from adult canine using density gradient separation method.The cMSCs attachment formed soon after seeding and grew into colonies with the appearance of fibroblastic cells.The osteogenic induction compound of Dexamethasone(Dex)、β-sodium glycerphosphate(β-GP)、ascorbic acid(AA)was added to passaged cMSCs and the proliferation and osteogenic differentiation of them was studied.The morphology of cells was observed by light micrograph and transmission electron microscope.The proliferation and growth characteristics of cMSCs were observed during primary and passage cultures through MTT.The differentiation were assayed by alkaline phophatase(ALP)and osteocalcin(OCN).We found the cMSCs have an active proliferative ability in primary and passage culture,and cMSCs under osteogenic induction have the typical characteristic of a secretory cell;the osteogenic induction compound may induce cMSCs to differentiate to osteoblasts.There are higher expression of ALP and OCN in passage 3 cMSCs under osteogenic induction than that of the osteoblasts osteogenic induction condition.Our research suggest the cMSCs in our culture system are mainly undifferentiated osteoprogenitors and can differentiate to osteoblast under osteogenic induction.更多还原