SD大鼠成骨细胞与兔骨髓间充质干细胞共育诱导成骨的实验研究

发布时间：2018-04-05 01:20

  本文选题：成骨细胞　切入点：骨髓间充质干细胞　出处：《辽宁医学院》2012年硕士论文

【摘要】：目的 通过应用Transwell培养板，将SD乳鼠颅盖骨成骨细胞与乳兔骨髓间充质干细胞（BMSCs）共培养，观察共培养环境下SD乳鼠颅盖骨成骨细胞对乳兔BMSCs向成骨细胞方向的诱导作用。 方法 取出生3天SD大鼠乳鼠颅盖骨，采用组织块培养法培养成骨细胞；取乳兔长骨，采用全骨髓培养法培养BMSCs，并用流式细胞仪分析细胞表面CD44、CD34、CD45、CD90的表达对其进行鉴定；应用Transwell双层细胞培养板将大鼠成骨细胞与兔骨髓间充质细胞共培养，实验组于培养上室接种成骨细胞，对照组培养上室不接种细胞，两组的培养下室均接种BMSCs。观察各组BMSCs形态的变化，，用酶标仪检测诱导分化后BMSCs的ALP活性，RT-PCR法分别检测两组BMSCs的骨钙素、骨形态发生蛋白-2（BMP-2）、Ⅰ型胶原基因的表达情况。 结果 1、大鼠成骨细胞与兔BMSCs共培养后，实验组BMSCs形态逐渐向成骨样细胞转化，而对照组无明显变化； 2、实验组BMSCs的ALP表达活性为0.365±0.016，对照组为0.17±0.012，即实验组BMSCs中ALP活性高于对照组。 3、实验组诱导后的BMSCs的骨钙素、BMP-2、Ⅰ型胶原基因有所表达，对照组无表达。 结论 乳鼠成骨细胞与乳兔BMSCs共培养后，能够诱导BMSCs向成骨细胞分化，BMSCs表达了成骨细胞的特异性表面标志。
[Abstract]:PurposeThe osteoblasts from calvaria of neonatal SD rats and bone marrow mesenchymal stem cells (BMSCs) of neonatal SD rats were co-cultured with Transwell culture plate to observe the induction of osteoblasts from calvaria to osteoblasts of newborn rabbits in co-cultured environment.MethodThe cranial calvaria of SD rats was taken out for 3 days and osteoblasts were cultured by tissue mass culture, and BMSCs were cultured by whole bone marrow culture method.Rat osteoblasts and rabbit bone marrow mesenchymal cells were co-cultured with Transwell double cell culture plate. The experimental group was inoculated with osteoblasts in the upper chamber, but the control group was not inoculated in the upper chamber.The morphological changes of BMSCs in each group were observed. The expression of osteocalcin, bone morphogenetic protein (BMP)-2 BMP-2 and type I collagen gene in BMSCs were detected by reverse transcription-polymerase chain reaction (RT-PCR) for detecting the ALP activity of BMSCs induced by differentiation.Result1. After co-culture of rat osteoblasts with rabbit BMSCs, the morphology of BMSCs in the experimental group was gradually transformed to osteoblast-like cells, while in the control group, there was no significant change.2. The ALP expression activity of BMSCs was 0.365 卤0.016 in the experimental group and 0.17 卤0.012 in the control group. That is, the ALP activity in the BMSCs of the experimental group was higher than that in the control group.3. Osteocalcin 2, type I collagen gene in BMSCs was expressed in the experimental group, but not in the control group.ConclusionAfter co-culture of neonatal rat osteoblasts and BMSCs, BMSCs could be induced to differentiate into osteoblasts and express the specific surface markers of osteoblasts.
【学位授予单位】：辽宁医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329

【参考文献】

相关期刊论文 前10条

1 马华梅;小儿生长和骨生化标志的改变[J];国外医学(儿科学分册);1999年02期

2 刘红,廖二元,伍贤平;骨钙素与代谢性骨病[J];国外医学.内分泌学分册;2004年04期

3 孟繁浩;熊志立;陶胜宇;李发美;;成骨细胞的生物学特征及其调控活性物质[J];化学与生物工程;2005年11期

4 刘宁;查振刚;王双利;林宏生;王国普;;成骨细胞诱导骨髓基质细胞体外成骨的初步研究[J];暨南大学学报(自然科学与医学版);2009年04期

5 马剑雄;马信龙;孙晓雷;骆巍;王志钢;李稚君;;体外培养成骨细胞对自体骨髓间充质干细胞定向成骨分化的影响[J];生物医学工程与临床;2009年04期

6 孟乘飞;张春礼;范宏斌;徐虎;白建萍;;EGF体外诱导骨髓基质干细胞增殖分化为成纤维细胞的实验研究[J];现代生物医学进展;2007年03期

7 林云锋;陈希哲;田卫东;阎征斌;郑晓辉;;大鼠脂肪源性血管内皮细胞的培养及其形态特征[J];生物医学工程学杂志;2006年04期

8 郝瑞锋;申长虹;;骨髓间充质干细胞向成骨细胞诱导的研究现状[J];天津医科大学学报;2007年01期

9 杨乃龙;林鹏;;骨髓间充质干细胞的分化潜能及临床应用价值[J];中国组织工程研究与临床康复;2007年03期

10 庄洪;徐无忌;;成骨细胞共育环境下大鼠骨髓间充质干细胞的成骨特性[J];中国组织工程研究与临床康复;2008年47期

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