人源单核细胞系28SC体外热原检查体系的可行性研究

杨泽岸; 陈晨; 郭龙静; 张媛; 吴彦霖

doi:10.13748/j.cnki.issn1007-7693.20240162

人源单核细胞系28SC体外热原检查体系的可行性研究

Feasibility Study of an in Vitro Pyrogen Detection System for Human-derived Monocyte Cell Line 28SC

摘要

摘要:
目的研究人源单核细胞系28SC体外检测热原的可行性。
方法将28SC分别与空白对照(IMDM)、脂多糖(lipopolysaccharide，LPS)共同孵育，以酶联免疫分析(ELISA)法检测孵育体系中相关细胞因子IL-6的含量变化。使用calcitriol刺激28SC，建立28SC热原检查体系；考察28SC体外检测热原体系最低检测限以及线性范围。
结果 calcitriol浓度为500 ng·mL⁻¹时，28SC-IL-1β、TNF-α法对LPS最低可检测到0.5 EU·mL⁻¹，对脂磷壁酸和酵母多糖(Zymosan)无反应，28SC-IL-6法对LPS最低可检测到0.125 EU·mL⁻¹。
结论本研究确定了28SC热原检查体系的活化物calcitriol浓度为500 ng·mL⁻¹，检测指标为细胞因子IL-6。

Abstract:
OBJECTIVE To investigate the feasibility of in vitro detection of pyrogen by the human-derived monocyte cell line 28SC.
METHODS 28SC was incubated separately with blank control(IMDM) and lipopolysaccharide(LPS), and the changes in the content of the relevant cytokine IL-6 in the incubation system were detected by enzyme-linked immunosorbent assay(ELISA). Stimulate 28SC with calcitriol to establish a 28SC pyrogen detection system, establishing the 28SC pyrogen detection system; investigating the minimum detection limit and linear range of the 28SC in vitro pyrogen detection system.
RESULTS When the concentration of calcitriol was 500 ng·mL⁻¹, the 28SC-IL-1β and TNF-α could detect 0.5 EU·mL⁻¹ for LPS, and did not react to LTA and Zymosan, and the 28SC-IL-6 could detect 0.125 EU·mL⁻¹ for LPS, and the detection limit of the 28SC pyrogen assay system was determined to be 0.5 EU·mL⁻¹.
CONCLUSION This study determines that the concentration of the activator calcitriol in the 28SC pyrogen detection system is 500 ng·mL⁻¹, and the detection indicator is cytokine IL-6.

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