狗骨髓間葉幹細胞之骨誘導分化潛能的體外研究

年齡增長後，骨髓中幹細胞(MSCs)數量明顯減少。另外，年齡增長後骨折癒合有明顯延遲情形，骨髓細胞骨生成能力和骨生成細胞增生明顯降低。臨床上，autologous bone雖然可以提供bone cell、骨誘導和應力支持，但graft量往往太少，形狀不吻合，無法完全吸收。異體移植則易造成免疫排斥和感染等問題。對年長病人而言，則可能需要足夠MSCs才能有效促進或加速骨缺損癒合。建立一個研究骨髓間葉幹細胞之大動物模式，以提供日後進行骨大缺損或骨折等骨組織缺損進行有效修復／再生相關研究之用，是必要的。因此，本研究旨在探討成年狗骨髓間葉幹細胞之骨組織細胞分化誘導及永質礦物化潛能。實驗結果發現狗脛骨骨髓間葉幹細胞經CD34篩選分離出CD34細胞進行培養。培養過程加入誘導因子，可見細胞形狀由梭狀變成類造骨細胞之立方形，並有顯著的鹼性磷酸酵素和第一型膠原蛋白表現。誘導15天時，可見造骨細胞外基質開始進行礦物化，並形成骨群體。本研究結果顯示狗骨髓CD34間葉幹細胞經分離、培養後，可被誘導分化成骨細胞／組織。

關鍵字

鹼性磷酸酵素；骨群體；間葉幹細胞；第一型膠原蛋白

並列摘要

With aging, the decrease in bone mass (or bone mineral density) could increase the occurrence of fracture. Thus, a decrease in personal bone mineral density could increase the occurrence of fracture during life. A fracture could result in loss of function or even death. Therefore, investigation on keeping bone tissue and cells functionally and prevention of damage of bone structure are imperative in regenerative medicine. There are significant decreases in bone marrow messenchymal stem cells (MSCs) and bone healing with aging. In addition, a delay in fracture healing and significant decrease in proliferation of osteoproganitor cells were noted. In order to establish a suitable big animal study model (using dog) for the study of differentiation potential and clinical application of bone marrow mesenchymal stem cells, the purpose of this study was to investigate the osteogenic potential of canine bone marrow mesenchymal stem cell after isolation and induction using density gradient separation method, CD markers and characterization of osteogenic differentiation phenotype expression. Seven and fifteen days after induction, appearance of cuboid-shaped osteoblasts and matrix mineralization were noted. It is concluded that canine marrow CD 34(superscript -) MSCs could be isolated, cultured and induced to differentiate into osteogenic lineages/tissues.

並列關鍵字

Alkine phosphatase activity ； bone colony ； mesenchymal stem cells ； type I collagen

國際替代計量

狗骨髓間葉幹細胞之骨誘導分化潛能的體外研究

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