组织工程与重建外科杂志

组织工程与重建外科杂志 ›› 2018, Vol. 14 ›› Issue (6): 311-313.doi: 10.3969/j.issn.1673-0364.2018.06.003

• 论著 • 上一篇    下一篇

大分子拥挤对脂肪干细胞增殖及其成骨分化能力的影响

耿英楠，韦敏，毛豪丽   

  1. 上海交通大学医学院附属第九人民医院
  • 收稿日期:2018-09-10 修回日期:2018-11-03 发布日期:2020-07-23
  • 作者简介:韦敏(E-mail:drweimin@163.com);毛豪丽(E-mail:maohaoli@126.com)

Efficacy of Macromolecule Crowding on the Proliferation and Osteogenic Differentiation of Adipose-Derived Stem Cells

GENG Yinnan,WEI Min,Mao Haoli   

  • Received:2018-09-10 Revised:2018-11-03 Published:2020-07-23

摘要： 目的 研究大分子拥挤环境对脂肪干细胞(ADSCs)增殖及体外成骨分化能力的影响。方法 将大分子拥挤试剂Ficoll 400和Ficoll 70分别以25 mg/mL和37.5 mg/mL的浓度混合,制备成50 mg/mL Fc混合液,用于体外培养ADSCs。通过CCK-8法、dsDNA核酸定量分析及BCA蛋白浓度测定,定量检测大分子拥挤环境对ADSCs增殖能力的影响;在不同培养时间对ADSCs进行成骨诱导,茜素红染色,检测ADSCs成骨分化能力。结果 CCK-8法检测显示大分子拥挤促进了ADSCs的增殖;dsDNA核酸定量分析及BCA蛋白浓度测定结果表明,大分子拥挤促进了ADSCs的DNA和蛋白质的合成;茜素红染色结果显示,大分子拥挤环境下ADSCs成骨分化能力增强,钙结节增加。结论 大分子拥挤环境对ADSCs的增殖及其体外成骨分化均有一定的促进作用。

关键词: 大分子拥挤, 成骨分化, 细胞外基质

Abstract: Objective To explore the effect of macromolecule crowding on the proliferation and osteogenic differentiation of adipose-derived stem cells(ADSCs).Methods The macromolecular crowders Ficoll 400 and Ficoll 70 were mixed with the concentrations of 25 mg/mL and 37.5 mg/mL to produce 50 mg/mL Fc mixture for the in vitro culture of ADSCs.CCK-8 test,dsDNA kit and BCA kit were used to quantitatively determine the effect of macromolecule crowding on the proliferation capacity of ADSCs under standard conditions.And the cells were subjected to osteogenic induction at different time points.Alizarin red staining was used to detect the changes in the osteogenic differentiation ability of ADSCs.Results CCK-8 test showed that the macromolecule crowding promoted the proliferation of stem cells.dsDNA analysis and BCA quantitative detection showed that the macromolecule crowding promoted the synthesis of ADSCs DNA and proteins.In addition,calcium nodules of the osteoblasts were enhanced showed by alizarin red staining,which proved that the osteoblastic differentiation ability was improved.Conclusion The macromolecular crowding niche promotes the osteogenic differentiation and proliferation of ADSCs in vitro.

Key words: Macromolecular crowding, Osteogenic differentiation, Extracellular matrix

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