Tang Yanjuan, Li Yi, Chen Huaiqing, Wu Qiaofeng, Yin Guangfu, Zhou Dali
Institute of Biomedical Engineering, West China Medical Center of Sichuan University, Chengdu 610041, China.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2006 Feb;23(1):142-6.
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), beta-sodium glycerphosphate (beta-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.
为研究培养的犬间充质干细胞（cMSCs）在成骨诱导下的生长和成骨特性。我们发现采用密度梯度分离法从成年犬中分离出cMSCs。接种后cMSCs很快附着并生长成集落，呈现成纤维细胞样外观。将地塞米松（Dex）、β-甘油磷酸钠（β-GP）、抗坏血酸（AA）组成的成骨诱导复合物添加到传代的cMSCs中，研究其增殖和成骨分化情况。通过光学显微镜和透射电子显微镜观察细胞形态。在原代培养和传代培养过程中通过MTT观察cMSCs的增殖和生长特性。通过碱性磷酸酶（ALP）和骨钙素（OCN）检测分化情况。我们发现cMSCs在原代培养和传代培养中具有活跃的增殖能力，成骨诱导下的cMSCs具有分泌细胞的典型特征；成骨诱导复合物可诱导cMSCs分化为成骨细胞。成骨诱导条件下第3代cMSCs中ALP和OCN的表达高于成骨细胞成骨诱导条件下的表达。我们的研究表明，我们培养体系中的cMSCs主要是未分化的骨祖细胞，在成骨诱导下可分化为成骨细胞。
