Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/13746
標題: 犬之間葉幹細胞之分離與特性
The Isolation and Characteristics of Canine Mesenchymal Stem Cells
作者: 簡國傑
Chien, Kuo-Chieh
關鍵字: dog

fracture
bone marrow
mesenchymal stem cells (MSCs)
platelet-rich fibrin (PRF)
osteogenesis
骨折
骨髓
間葉幹細胞
富含血小板纖維
骨化作用
出版社: 獸醫學系暨研究所
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摘要: 間葉幹細胞(mesenchymal stem cell)是骨髓中重要的成分之一,由於間葉幹細胞擁有高增殖力以及多潛能的分化能力,如成骨化、脂肪化及軟骨化等,故近二十年來間葉幹細胞遂成為細胞治療的工具。然而有關犬間葉幹細胞的功能及分化能力之研究甚少。本實驗之目的為先觀察犬隻間葉幹細胞,於體外培養的增殖狀況及成骨化的情形,並利用共同培養的方式,分析富含血小板纖維(platelet-rich fibrin)對間葉幹細胞之影響。富含血小板纖維的製備,由不加抗凝劑的週邊血液,直接離心而得。內含有很多生長因子,主要為轉化成長因子-β (transforming growth factors-β) 、血小板源生長因子(platelet-derived growth factor)及類胰島素生長因子(insulin-like growth factor)。犬骨髓樣本的取得,由2007至2009年,從中興大學獸醫教學醫院之骨折修復手術中採集,共23株。經體外培養後,可見其表現間葉幹細胞之相關蛋白,如間葉細胞骨架蛋白(vimentin)及細胞黏附蛋白(fibronectin)。除此之外,利用專一性細胞表面蛋白, CD44陽性及CD34陰性區別出犬間葉幹細胞。體外培養的犬間葉幹細胞在第一次繼代到第三次繼代表現出高增殖力。並且在培養液中添加皮質類固醇(dexamethasone)、β-磷酸甘油(β-glycerolphosphate)及抗壞血酸(ascorbic acid)誘導成骨化。三週後,可見細胞型態轉為多角狀(polygonal appearance)、生長成叢狀(clump formation)及細胞鹼性磷酸脢(alkaline Phosphatase)活性上升。將犬間葉幹細胞與富含血小板纖維共同培養在不同營養環境的培養液中,可知富含血小板纖維具有促進犬間葉細胞增殖的能力,但無法促進成骨分化。
Mesenchymal stem cells (MSCs) are important composition in bone marrow and can be a promising tool for cytotherapy. MSCs possess highly proliferative and multi-potent differentiation for osteogenesis, adipogenesis and chondrogenesis. However, the function and further differential pattern of canine MSCs still remain unclear. The aim of this study was to investigate proliferation and osteogenesis of canine MSCs co-cultured with platelet-rich fibrin (PRF). PRF is a concentration of platelets from peripheral blood without anticoagulant. PRF contains growth factors including transforming growth factor β, platelet-derived growth factor and insulin-like growth factor. Canine MSCs were obtained from bone marrows of 23 patients referred to our hospital for bone fracture repair between 2007 and 2009. They showed positive immunoreaction with vimentin, fibronectin, and specific MSCs surface markers CD44 and negative immunoreaction with CD34. Canine MSCs showed high proliferation from passage first to passage third. Canine MSCs were induced to develop osteogenesis by the medium containing dexamethason, β-glycerophosphate and, ascorbic acid and the morphology was found to be transformed into polygonal appearance, showed clumps formation, and increased the alkaline phosphatase activity during 3 weeks. Canine MSCs were co-cultured with equal volume of PRF with 2.5%, 5.0%, and 10% of fetal bovine serum. PRF promoted proliferation of canine MSCs and did not promote the osteogenic differentiation in vitro.
URI: http://hdl.handle.net/11455/13746
其他識別: U0005-1007200914292400
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-1007200914292400
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