从干细胞和血清中特征细胞外囊泡的局限性和挑战
Sara Escudero-Cernuda1, Noemi Eiro2, María Fraile3
1Department of Physical and Analytical Chemistry, University of Oviedo, Avda. Julián Clavería, 8, 33006, Oviedo, Asturias, Spain.
Mikrochimica acta
|April 21, 2025
概括
对无细胞疗法至关重要的外体体进行表征是具有挑战性的,因为方法不一致. 这项研究引入了用于纯度评估的尺寸排除色谱,揭示了商业外体的低纯度,并使粒子度估计成为可能.
科学领域:
- 生物技术是生物技术.
- 细胞生物学 细胞生物学
- 纳米医学是一种纳米医学.
背景情况:
- 外体是纳米尺寸的细胞外囊泡,对于细胞间通信至关重要.
- 它们对再生医学,免疫疗法和药物输送具有前景.
- 缺乏标准化的表征方法,阻碍了临床翻译.
研究的目的:
- 描述人类子宫介质干细胞 (hUCESC) 衍生的细胞外囊泡 (EV).
- 评估EV表征方法的当前局限性.
- 为EV纯度评估引入和验证大小排除高性能液体染色学 (SE-HPLC).
主要方法:
- 使用超离心法将电动汽车与hUCESC隔离.
- 使用总蛋白质测定和SE-HPLC评估EV纯度.
- 从脂肪干细胞和人血清中获得的hUCESC-EV和商业EV之间的纯度比较.
主要成果:
- 总蛋白质测定容易受到污染,因此需要多种方法来准确确定囊泡蛋白质.
- 电子电源的起源影响纯度,复杂的介质导致更大的干扰.
- SE-HPLC在商业外体中显示出较低的纯度,强调了报告蛋白质和脂质纯度的必要性.
- 将SE-HPLC与蛋白质试验相结合,可以根据囊泡蛋白质度估计颗粒度.
结论:
- 标准化和验证的方法对于可靠的外体表征是必不可少的.
- SE-HPLC是评估EV纯度的一个有价值的工具,可以与蛋白质测定集成.
- 商业外体产品需要对蛋白质和脂质纯度进行严格的质量控制.
相关概念视频
01:20Overview Of Cell Separation And Isolation
Cell separation was first achieved in 1964 by S. H. Seal, who separated large tumor cells from the smaller blood cells using filtration. Two years later, Pohl and Hawk performed experiments on how cells respond differently to a nonuniform electric field based on the cell type. Such observations were the inception of cell separation methods, which allow isolating a single cell type from a heterogeneous sample.
01:23Flow Cytometry
The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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