中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (31): 4935-4939.doi: 10.12307/2022.732

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

浓缩生长因子干预大鼠骨髓间充质干细胞的增殖、成骨分化及迁移

汤 凯1,2,招文华1,2,尚 奇1,2,沈耿杨2,3,张志达2,3,何嘉辉1,2,张 鹏1,2,余富勇1,2,陈桂锋1,2, 任 辉2,3,江晓兵2,3,余 翔2,3   

  1. 1广州中医药大学,广东省广州市   510405;2广州中医药大学岭南医学研究中心,广东省广州市   510405;3广州中医药大学第一附属医院,广东省广州市   510405
  • 收稿日期:2021-11-04 接受日期:2021-12-10 出版日期:2022-11-08 发布日期:2022-04-24
  • 通讯作者: 余翔,博士,主治医师,广州中医药大学岭南医学研究中心,广东省广州市 510405;广州中医药大学第一附属医院,广东省广州市 510405
  • 作者简介:汤凯,男,1994年生,江西省丰城市人,汉族,广州中医药大学在读硕士,主要从事骨质疏松、骨组织工程基础与临床研究。
  • 基金资助:
    国家自然科学基金面上项目(81774338),项目负责人:江晓兵;广东省医学科研基金项目(A2021320),项目负责人:余翔;广州中医药大学青年培育项目(2019QNPY04),项目负责人:余翔

Effects of concentrated growth factors on the proliferation, osteogenic differentiation and migration of rat bone marrow mesenchymal stem cells

Tang Kai1, 2, Zhao Wenhua1, 2, Shang Qi1, 2, Shen Gengyang2, 3, Zhang Zhida2, 3, He Jiahui1, 2, Zhang Peng1, 2, Yu Fuyong1, 2, Chen Guifeng1, 2, Ren Hui2, 3, Jiang Xiaobing2, 3, Yu Xiang2, 3   

  1. 1Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China; 2Lingnan Medical Research Center, Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China; 3First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China
  • Received:2021-11-04 Accepted:2021-12-10 Online:2022-11-08 Published:2022-04-24
  • Contact: Yu Xiang, MD, Attending physician, Lingnan Medical Research Center, Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China; First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China
  • About author:Tang Kai, Master candidate, Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China; Lingnan Medical Research Center, Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China
  • Supported by:
    General Program of National Natural Science Foundation of China, No. 81774338 (to JXB); Medical Research Foundation of Guangdong Province, No. A2021320 (to YX); Youth Training Program of Guangzhou University of Chinese Medicine, No. 2019QNPY04 (to YX)

摘要:

文题释义:
浓缩生长因子:不含任何添加剂离心出来的一种高度浓缩生长因子的血纤维蛋白,又称血液中的“不老因子”,白细胞、血小板及各类生长因子是它的主要成分,如血管内皮生长因子、成纤维细胞生长因子等,具有组织再生、炎症调节、抗感染等特性,在骨科、口腔、美容等领域应用广泛。
细胞迁移:指细胞从一个地方移动到另一个地方的复杂而异构的过程,分为单细胞迁移和集体细胞迁移。单细胞迁移分伪足运动和间叶细胞运动,伪足运动的特征是细胞以圆形椭球体的形式运动,没有局部黏附和细胞附着,但有推进泡的帮助;间叶细胞运动则包括细胞外基质的强烈局部附着、细胞骨架的收缩等。集体细胞迁移主要特征是细胞队列通过细胞外基质运动,并保存功能细胞-细胞连接。

背景:在研究骨髓间充质干细胞的增殖、成骨分化基础上,揭示其迁移机制是再生医学研究的热门课题之一。
目的:探究浓缩生长因子对大鼠骨髓间充质干细胞增殖、成骨分化及迁移能力的影响。
方法:大鼠心脏静脉取血,变速离心提取浓缩生长因子。体外培养大鼠骨髓间充质干细胞,分为实验组和对照组,实验组分别用含体积分数5%,10%,20%浓缩生长因子的α-MEM完全培养基进行培养,对照组用不含浓缩生长因子的α-MEM完全培养基进行培养。应用CCK-8法检测细胞增殖能力,划痕实验、Transwell迁移小室实验检测细胞迁移能力,茜素红染色检测细胞成骨分化能力。
结果与结论:①不同体积分数的浓缩生长因子均能促进大鼠骨髓间充质干细胞增殖;②划痕实验显示实验组均能够促进大鼠骨髓间充质干细胞迁移,其中12 h时体积分数10%浓缩生长因子组迁移速度最快;Transwell迁移小室实验结果显示,实验组大鼠骨髓间充质干细胞迁移数目均高于对照组,且体积分数10%浓缩生长因子组的细胞迁移数量更多;③茜素红染色显示,实验组在7 d时钙结节数量均多于对照组;④结果表明,不同体积分数的浓缩生长因子均能够促进大鼠骨髓间充质干细胞的体外增殖、分化及迁移,其中体积分数10%浓缩生长因子具有较快的促进细胞迁移能力。

https://orcid.org/0000-0001-9181-9692 (余翔) 

中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程

关键词: 浓缩生长因子, 骨髓间充质干细胞, 细胞增殖, 成骨分化, 细胞迁移

Abstract: BACKGROUND: On the basis of studying the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells, it is one of the hot topics in regenerative medicine to reveal their migration mechanism.  
OBJECTIVE: To investigate the effects of concentrated growth factor on proliferation, osteogenic differentiation, and migration of rat bone marrow mesenchymal stem cells.
METHODS:  Concentrated growth factor was extracted from rat heart vein by variable speed centrifugation. Rat bone marrow mesenchymal stem cells were cultured in vitro and divided into experimental and control groups. Cells in the experimental groups were cultured in α-MEM complete medium supplemented with 5%, 10% and 20% concentrated growth factors by volume fraction, separately. Cells in the control group were cultured in α-MEM complete medium without concentrated growth factor. Cell proliferation was detected by CCK-8 assay. Cell migration was detected by scratch test and Transwell migration chamber assay. Osteogenic differentiation was detected by alizarin red staining.  
RESULTS AND CONCLUSION: (1) Concentrated growth factor with different volume fractions could promote the proliferation of rat bone marrow mesenchymal stem cells with different volume fractions. (2) The scratch test showed that the migration of rat bone marrow mesenchymal stem cells could be promoted in the experimental groups, and the migration rate of 10% concentrated growth factor group was fastest at 12 hours. Transwell migration chamber assay showed that the number of migrated rat bone marrow mesenchymal stem cells in the experimental groups was higher than that in the control group. The number of migrated rat bone marrow mesenchymal stem cells was highest in the 10% concentrated growth factor group. (3) Alizarin red staining showed that the total number of calcium nodules in the experimental groups was higher than that in the control group on day 7. (4) The results showed that all concentrated growth factors with different volume fractions could promote the proliferation, differentiation, and migration of rat bone marrow mesenchymal stem cells in vitro. The 10% concentrated growth factor had a faster ability to promote the migration of rat bone marrow mesenchymal stem cells.

Key words: concentrated growth factors, rat bone marrow mesenchymal stem cells, cell proliferation, osteogenic differentiation, cell migration

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