组织工程与重建外科杂志

组织工程与重建外科杂志 ›› 2013, Vol. 9 ›› Issue (3): 121-124.doi: 10.3969/j.issn.1673-0364.2013.03.001

• 论著 •    下一篇

不同血清浓度对淋巴管内皮细胞培养的影响

葛敏，蒋朝华，戴婷婷，程佳，李圣利   

  1. 上海交通大学医学院附属第九人民医院整复外科
  • 发布日期:2020-07-23

The Effects of Serum Concentration on the Culture of Lymphatic Endothelial Cells

GE Min, JIANG Zhaohua, DAI Tingting, CHENG Jia, LI Shengli   

  1. Department of Plastic and Reconstructive Surgery, Shanghai Ninth People’s Hospital, Shanhai Jiaotong University School of Medicine, Shanghai 200011, China.
  • Published:2020-07-23
  • Contact: 上海交通大学科技创新发展专项资金—理工交叉研究基金

摘要： 目的探讨不同浓度的血清在人淋巴管内皮细胞培养中的作用,得出最优血清浓度,优化淋巴管内皮细胞(Lymphatic endothelial cells,LECs)的培养方案。方法通过免疫磁珠法获得人LECs,将不同浓度(0%、2.5%、5%、7.5%、10%)的胎牛血清加入EGM-2培养液中,培养LECs,并通过免疫荧光、RT-PCR、流式分析、成管试验、吞脂试验等方法,鉴定LECs特异性标记及功能。结果通过免疫磁珠法获得的人LECs表达特异性指征和功能。不同浓度的血清培养液对人LECs的吞脂功能无明显影响,无血清组丧失淋巴管成管能力,7.5%血清组成管能力和增殖能力最强。结论人LECs培养过程中,血清在细胞增殖和功能保持方面具有重要作用,最适宜的血清浓度是7.5%,此浓度血清培养的细胞其增殖能力以及功能都是最优的。

关键词: 血清, 淋巴管内皮细胞, 培养

Abstract: Objective To explore the effects of serum concentration on the culture of Lymphatic endothelial cells（LECs） and determine an optimal serum concentration on the culture of LECs.Methods LECs were isolated and identified by immunomagnetic beads sorting method,then divided into 5 experimental groups and cultured with 5 different serum concentrations of EGM-2 respectively（0%,2.5%,5%,7.5% and 10%）.The proliferation and function of the LECs were examined and compared by immuno-fluoresence staining,RT-PCR,flow cytometry,matrigel assay and Dil-Ac-LDL uptake experiment.Results The cells obtained by immunomagnetic beads sorting method demonstrated the specific marks and functions of LECs.There were no significant differences on Dil-Ac-LDL uptake of LECs in each group.The tube-like structure was not found in the 0% serum concentration group.It is revealed that the lymphatic vessels formation and proliferation were the highest in 7.5% serum concentration group.Conclusion The serum plays an important role in LECs culture,especially in cell proliferation and function.The optimal serum concentration is 7.5% serum concentration,which is conducive to the proliferation and function of LECs.

Key words: Serum, Lymphatic endothelial cells, Culture

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