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碱性成纤维细胞生长因子对人骨髓间充质干细胞 HLA-DR 表达的影响

Effect of bFGF on HLA-DR expression of human bone marrow-derived mesenchymal stem cells.

作者信息

Dighe Pratiksha A, Viswanathan Pachaiyappan, Mruthunjaya Ashwin K, Seetharam Raviraja N

机构信息

Stempeutics Research, Shirdi Sai Baba Cancer Hospital, Manipal, India.

出版信息

J Stem Cells. 2013;8(1):43-57.

Abstract

There has been a steady rise in the therapeutic applications of bone marrow mesenchymal stem cells (BM-MSCs) because of their unique properties of multilineage differentiation and immune modulation as well as the ease in isolation. However, up-regulation of surface HLA-DR levels when maintaining MSCs in culture under the influence of mitotic factors such as Basic fibroblast growth factor (bFGF) is an area of concern when considering them for the purpose of clinical applications. Thus, we investigated the association of bFGF supplemented to the culture media and the surface expression levels of HLA-DR in BM-MSCs in order to optimize the yield, while keeping HLA-DR levels under permissible levels. Human BM-MSCs were culture expanded in the absence of bFGF and in the presence of 1 ng/ml or 2 ng/ml bFGF. The HLA-DR profile of the cultures was analyzed at the end of each passage. On comparing the percent HLA-DR+ cell population at different concentrations as well as absence of bFGF, significant differences were not observed in the HLA-DR expression levels of the MSC cultures which had reached complete confluence. However, variations in HLA-DR expressions levels were seen which could be traced to the age of cells in culture with values drastically reduced to below 4% on maintaining MSCs typically two to three days beyond achieving full confluence. On the basis of the findings from this study, no significant correlation could be established on the effect of bFGF in modulating HLA-DR surface expression of BM-MSCs. Instead, the data are suggestive of the reasoning that the duration for which BM-MSCs are maintained in culture directly influences their phenotypic characteristics in terms of HLA-DR expression levels, with lowest levels achieved on their prolonged maintenance in culture.

摘要

由于骨髓间充质干细胞(BM-MSCs)具有多谱系分化和免疫调节的独特特性以及易于分离,其治疗应用一直在稳步增加。然而,在诸如碱性成纤维细胞生长因子(bFGF)等有丝分裂因子的影响下,在培养中维持间充质干细胞时表面HLA-DR水平的上调是在考虑将其用于临床应用时需要关注的一个领域。因此,我们研究了添加到培养基中的bFGF与BM-MSCs中HLA-DR表面表达水平之间的关联,以便在将HLA-DR水平保持在允许水平的同时优化产量。人BM-MSCs在无bFGF以及存在1 ng/ml或2 ng/ml bFGF的情况下进行培养扩增。在每次传代结束时分析培养物的HLA-DR谱。在比较不同浓度以及无bFGF时HLA-DR+细胞群体的百分比时,在已达到完全汇合的MSC培养物的HLA-DR表达水平上未观察到显著差异。然而,观察到HLA-DR表达水平存在变化,这可以追溯到培养细胞的年龄,在达到完全汇合后通常再维持两到三天的MSCs中,其值急剧降至4%以下。基于本研究的结果,无法确定bFGF对调节BM-MSCs的HLA-DR表面表达的影响之间存在显著相关性。相反,数据表明这样一种推断,即BM-MSCs在培养中维持的持续时间直接影响其在HLA-DR表达水平方面的表型特征,在培养中延长维持时间时达到最低水平。

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