本文關鍵詞：BrdU標記對大鼠骨髓間充質(zhì)干細胞增殖與成骨能力的影響　出處：《福建醫(yī)科大學》2010年碩士論文　論文類型：學位論文

  更多相關文章： BrdU 骨髓間充質(zhì)干細胞 組織工程

【摘要】： 目的:觀察5-溴-2-脫氧尿嘧啶核苷(5-bromo-2-deoxyuridine, BrdU)標記前后對SD大鼠骨髓間充質(zhì)干細胞(Mesenchymal stem cells, MSCs)增殖與成骨能力的影響。 方法:直接貼壁法分離培養(yǎng)大鼠骨髓MSCs, 0.2%Brdu標記后檢測其增殖能力、克隆形成能力;經(jīng)成骨誘導液誘導培養(yǎng)3周后,利用茜素紅染色,熒光免疫組化和Real Time PCR觀測標記前后MSCs的成骨能力的變化情況。 結(jié)果:BrdU標記后MSCs克隆形成能力明顯減弱,在克隆個數(shù)和面積上均小于未標記組(P0.05);在增殖能力方面, BrdU標記后第4 d開始進入對數(shù)增長期時增殖較慢,其中第6 d,8 d,12 d和14 d兩組之間有顯著性差別,且BrdU標記組增殖高峰較低(P0.05);在成骨能力方面,經(jīng)成骨誘導后標記組與未標記組的MSCs都表達骨鈣素、I型膠原,并且都有明顯的鈣結(jié)節(jié)形成,Real Time PCR檢測顯示標記前后MSCs在骨鈣素基因的表達方面無顯著性差異(P0.05)。 結(jié)論:BrdU標記抑制MSCs的克隆增殖形成能力,但不會降低其成骨分化能力,可以廣泛應用于MSCs作為種子細胞的干細胞研究的示蹤劑。
[Abstract]:Objective: to observe 5-bromo-2-deoxyuridine (5-bromo-2-deoxyuridine). Effects of BrdU labeling on the proliferation and osteogenesis of bone marrow mesenchymal stem cells (MSCs) in SD rats. Methods: MSCs were isolated from rat bone marrow by direct adherent method. The proliferative ability and clone formation ability were detected after labeling with 0.2 Brdu. The osteogenic ability of MSCs was observed by alizarin red staining, fluorescence immunohistochemistry and Real Time PCR. Results the clone forming ability of MSCs was obviously weakened after being labeled with 1: BrdU, and the number and area of clone were lower than that of the unlabeled group (P0.05). In terms of proliferative ability, the proliferation was slower at the beginning of logarithmic growth at the 4th day after BrdU labeling, and there was a significant difference between the two groups on the 6th day, 8 days and 14 days. The peak of proliferation in BrdU labeled group was lower than that of P0.05; In terms of osteogenic ability, osteocalcin type I collagen was expressed in MSCs of both the labeled group and the unlabeled group after osteogenic induction, and obvious calcium nodules were formed. Real Time PCR showed that there was no significant difference in the expression of osteocalcin gene in MSCs before and after labeling. Conclusion the MSCs labeled with 1: BrdU inhibits the ability of clone proliferation but does not decrease its osteogenic differentiation. It can be widely used as a tracer for stem cell research of seed cells by using MSCs as a tracer.
【學位授予單位】：福建醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R329

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