Initial characterisation of exosomes released by umbilical cord-derivedmesenchymal stem cells and mature dendritic cells, under ‘Good Manufacturing Practice’ conditions

Hoang Huong Diem, Bui Thi Van Khanh, H. Nhung, N. Liêm, T. Uyen
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Abstract

Exosomes represent an important mode of intercellular communication and play key roles in many physiological and pathological processes. Exosomes have hitherto exhibited their capacity to modulate biological activities through their carrying of functional molecules such as proteins, lipids, and genetic materials. In the current study, we investigated exosomes released by mature dendritic cells (mDCs) and umbilical cord-derived mesenchymal stem cells (UCMSCs) under Good Manufacturing Practice (GMP) conditions. Ultracentrifugation was used to isolate and purify the exosomes. Additionally, a transmission electron microscope (TEM) and immunoblotting were used to characterise exosomal morphology and markers. The preliminary results showed that both mDCs and UCMSCs secreted exosomes into GMP culture media. Exosomes exhibited a cup-shaped morphology and showed positive for CD63. Additionally, no difference was observed between mDC-derived exosomes and UCMSC-derived exosomes regarding marker expression or morphology. This data indicates the potential for further development of GMP exosomes for clinical application.
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脐带源性间充质干细胞和成熟树突状细胞在“良好生产规范”条件下释放外泌体的初步表征
外泌体是细胞间通讯的重要方式,在许多生理和病理过程中起着关键作用。迄今为止,外泌体通过携带功能分子(如蛋白质、脂质和遗传物质)显示出调节生物活动的能力。在本研究中,我们研究了成熟树突状细胞(mDCs)和脐带源性间充质干细胞(UCMSCs)在良好生产规范(GMP)条件下释放的外泌体。采用超离心分离纯化外泌体。此外,透射电子显微镜(TEM)和免疫印迹法用于表征外泌体形态和标记物。初步结果表明,mDCs和UCMSCs均在GMP培养基中分泌外泌体。外泌体呈杯状,CD63阳性。此外,mdc衍生的外泌体和ucmsc衍生的外泌体在标记物表达或形态上没有差异。这一数据表明GMP外泌体有进一步开发用于临床应用的潜力。
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