探討活血化瘀藥物對間質幹細胞存活，更新，多能分化性之影響

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Title:	探討活血化瘀藥物對間質幹細胞存活，更新，多能分化性之影響 Effects of Promoting Blood Circulation compounds on survival self renewal and multilineage differentiation of mesenchymal stem cells
Authors:	劉玫孜;Mei-Tzu Liu
Contributors:	醫學檢驗生物技術學系碩士班
Keywords:	間質幹細胞;丹參酚酸;脂肪分化;mesenchymal stem cell;salvianolic acid b;adipogenisis
Date:	2013-08-22
Issue Date:	2013-10-02 11:27:26 (UTC+8)
Publisher:	中國醫藥大學
Abstract:	人類臍帶血間質幹細胞(mesenchymal stem cells)擁有在體外分化成骨細胞、脂肪細胞及血管內皮細胞的能力，並可當作於細胞療法或組織工程中的細胞來源。本研究中將間質幹細胞分化為骨細胞、脂肪細胞以及血管內皮細胞，並且探討丹參酚酸(salvianolic acid B, Sal B)對於間質幹細胞之存活率、自我更新以及分化能力的影響。研究首先藉由流式細胞儀鑑定間質幹細胞特性，可表現特異性蛋白包括CD90、CD44、CD146及STRO-1 等。其次，於不同培養環境誘導下，成功將間質幹細胞誘導分化為骨細胞、脂肪細胞以及血管內皮細胞，分化後細胞之p53及p65表現下降，藉由化學染色證實間質幹細胞分別分化成脂肪細胞及骨細胞。間質幹細胞可在誘導分化成血管內皮細胞，藉由流式細胞儀發現細胞表面內皮細胞特異性蛋白包括CD31、TM及VE-Cadherin，並藉由血管新生試驗發現，分化而成的類血管內皮細胞可形成網狀的血管結構。另一部份實驗結果也發現經丹參酚酸作用之後間質幹細胞的增生能力受抑制，而來自不同個體檢體之臍帶血間質幹細胞的分化中，發現骨分化、脂肪分化能力存在差異，不同來源之間質幹細胞分化能力上也有差異。 Human cord blood-derived mesenchymal stem cells (MSCs) have the potential to differentiate into mesenchymal tissues like osteocytes, adipocytes and endothelial cells in vitro, thus can be useful as a source of cells for cell therapy or tissue engineering. In this study, MSCs were induced to differentiate into osteocytes, adipocytes and endothelial cells,and we alsostudy theeffects of salvianolic acid B(Sal B)on survival, self-renewal, and differentiation of mesenchymal stem cells. Byusingflow cytometry,MSCs were characterizedpositive of the specific markers CD90, CD44, CD146 and STRO-1. MSCswere successfully differentiated into adipocytes and osteocytes after cultivation in condition media. In addition, the expression of p53 and p65 were decreased after MSCs differention. MSCs could be induced into endothelial-like cells, which could express the specific markers of CD31, TM and VE-cadherin. Matrigel angiogenesis assay displayed that endothelial differentiated cellswere able to form the capillary network.The proliferation ability of MSCs were inhibited by Sal B. MSCs isolated from human bone marrow (BM-MSC) or cord blood from different donors(clone 10-14) exhibited quantitative differences in their ability to differtiated into adipocyte.
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