Laboratory of Parasitology, General Karol Kaczkowski Military Institute of Hygiene and Epidemiology, Warsaw, Poland.
Laboratory of Parasitology, General Karol Kaczkowski Military Institute of Hygiene and Epidemiology, Warsaw, Poland
In Vivo. 2021 Jan-Feb;35(1):283-289. doi: 10.21873/invivo.12257.
BACKGROUND/AIM: Functional and quantitative Treg cell defects have been identified in a variety of autoimmune diseases. Therefore, Tregs are a major pharmaceutical target for these disorders. In the last decades, studies have been mainly focused on the identification and experimental understanding of the activity of Tregs and their mechanisms of action.
This study describes how overnight storage of isolated peripheral blood mononuclear cells in different media (PBS pH 7.3, PBS pH 7.3 containing 0.5% BSA, RPMI 1640 and RPMI 1640 containing 10% FBS) affects the viability and expression of the commonly used markers for Tregs identification: CD25, CD127, CTLA-4, GITR, PD-1, FoxP3 and Helios.
Incorrectly selected storage conditions (temperature, time, medium) may affect the expression of surface and intracellular markers, thus, compromising the quality of the obtained results.
Appropriate protocols of cell isolation and storage are important for providing appropriate conditions for cell growth. This is crucial when analyzing small cell populations like Tregs.
背景/目的：在多种自身免疫性疾病中，已经鉴定出功能性和定量的调节性 T 细胞缺陷。因此，Tregs 是这些疾病的主要药物靶点。在过去的几十年中，研究主要集中在鉴定和实验理解 Tregs 的活性及其作用机制上。
本研究描述了分离的外周血单核细胞在不同培养基（PBS pH 7.3、含 0.5%BSA 的 PBS pH 7.3、RPMI 1640 和含 10%FBS 的 RPMI 1640）中过夜储存如何影响常用于鉴定 Tregs 的标志物的表达：CD25、CD127、CTLA-4、GITR、PD-1、FoxP3 和 Helios。
不正确选择的储存条件（温度、时间、培养基）可能会影响表面和细胞内标志物的表达，从而影响获得结果的质量。
适当的细胞分离和储存方案对于为细胞生长提供适当的条件很重要。当分析像 Tregs 这样的小细胞群时，这一点至关重要。
