Protein contaminations impact quantification and functional analysis of extracellular vesicle preparations from mesenchymal stromal cells.

IF 1.1 Q4 CELL & TISSUE ENGINEERING Journal of Stem Cells & Regenerative Medicine Pub Date : 2015-12-31 eCollection Date: 2015-01-01
Meaghan Stolk, Martina Seifert
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引用次数: 0

Abstract

Extracellular vesicles (EV) awakened interest in the research on mesenchymal stromal cells by exploring their paracrine effects. However, many isolation protocols use bovine serum albumin (BSA) during the preparation of the EV. Therefore, we produced 'sham' BSA-EV and tested them in comparison to EV derived from mesenchymal stromal cells (MSC-EV). We found that BSA-EV did not express MSC-specific surface markers like CD29 and CD90. However, they were capable of reducing serum-starvation induced apoptosis in vitro in a kidney epithelial cell line to a similar extent as MSC-EV as measured by Annexin V/7-Aminoactinomycin D labeling and flow cytometry.

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蛋白质污染影响间充质间质细胞胞外囊泡制备的定量和功能分析。
细胞外囊泡(Extracellular vesicles, EV)通过探索间充质间质细胞的旁分泌作用,唤醒了人们对间充质间质细胞研究的兴趣。然而,许多分离方案在制备EV时使用牛血清白蛋白(BSA)。因此,我们制造了“假”BSA-EV,并将其与间充质间质细胞(MSC-EV)衍生的EV进行了比较。我们发现BSA-EV不表达msc特异性表面标记物如CD29和CD90。然而,通过Annexin V/7-氨基放线菌素D标记和流式细胞术检测,它们能够在体外减少血清饥饿诱导的肾上皮细胞系的凋亡,其程度与MSC-EV相似。
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来源期刊
CiteScore
3.40
自引率
0.00%
发文量
5
审稿时长
14 weeks
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