免疫磁珠法体外促脐带血调节性T细胞增殖能力的研究*

doi:10.3969/j.issn.1671-2587.2020.05.020

临床输血与检验 ›› 2020, Vol. 22 ›› Issue (5): 526-530.DOI: 10.3969/j.issn.1671-2587.2020.05.020

免疫磁珠法体外促脐带血调节性T细胞增殖能力的研究^*

苏木雅, 杜华, 师迎旭

010050 内蒙古医科大学（苏木雅,杜华）;
内蒙古医科大学附属医院（师迎旭）

收稿日期:2019-11-24 出版日期:2020-10-20 发布日期:2020-10-16
通讯作者: 师迎旭,男,博士,副研究员,主要从事细胞生物学方面研究,（E-mail）shiyingxu@126.com。
作者简介:苏木雅（1997-）,女,蒙古族,硕士研究生,主要从事分子诊断方面研究,（E-mail）378950474@qq.com。
基金资助:
*本课题受内蒙古自治区自然科学基金（No.2019LH080-32）、内蒙古自治区高等学校青年科技英才支持计划（No.NJYT-18-B18）、内蒙古医科大学科技百万工程（No.YKD20-17KJBW004）、内蒙古医科大学附属医院一般科研项目（No.NYFY YB040）资助

In Vitro Expansion of Human Umbilical Cord Blood Regulatory T Cells by Magnetic Beads

SU Mu-ya, DU Hua, SHI Ying-xu

The Affiliated Hospital of Inner Mongolia Medical University, Huhehaote 010050

Received:2019-11-24 Online:2020-10-20 Published:2020-10-16

摘要/Abstract

摘要： 目的分离人脐带血CD4⁺CD25⁺调节性T细胞（regulatory T cells, Tregs）,通过扩增磁珠进行体外扩增培养并检测其功能。方法利用分离磁珠分离人脐带血中的CD4⁺CD25⁺Tregs。将分离获得的CD4⁺CD25⁺Tregs与扩增磁珠共同培养。通过淋巴细胞分析实验观察脐血中淋巴细胞的含量,通过磁珠分选观察脐血中调节性T细胞含量,并通过PI染色观察分选到的细胞的活力。通过不同的培养条件（磁珠组、对照组）观察免疫磁珠对于Tregs的扩增能力。结果脐血中CD4⁺细胞中CD4⁺CD25⁺ Tregs含量约为4.5%,CD4⁺CD25^-细胞含量为95.5%。经过分离磁珠分选后可见Tregs被有效富集,PI染色检测细胞活性无损失,含量约为81%。与人T细胞扩增磁珠共培养后,可见Tregs被有效扩增。结论免疫分离磁珠能够有效的富集脐血单个核中的Tregs,且无明显的损伤。通过实验可见T细胞扩增磁珠确实能够很有效的在体外扩增调节性T细胞,但其生物学能力的评估仍需要进一步的研究证实。

关键词: 调节性T细胞, 扩增磁珠

Abstract: Objective To isolate CD4⁺CD25⁺ regulatory T cells (Tregs) from human umbilical cord blood (UCB), and to amplify and culture in vitro CD4⁺CD25⁺Tregs by immunomagnetic beads and report these function. Methods CD4⁺CD25⁺ Tregs were isolated from human UCB by immunomagnetic separation beads. The isolated CD4⁺CD25⁺ Tregs were co-cultured with expanded immunomagnetic beads. The content of lymphocyte in UBC was observed by lymphocyte analysis experiment, Tregs by magnetic beads sorting, and selected cell viability by PI staining. The proliferation ability of immunomagnetic beads to Tregs was observed under different culture conditions (immunomagnetic beads group, control group). Results CD4⁺CD25⁺Tregs and CD4⁺CD25^-cells in CD4⁺UCB were about 4.5% and 95.5%, respectively. Tregs were effectively enriched by magnetic beads sorting and cells stained with PI showed no loss of cell viability, and the content was about 81%. After co-culture with human T cell amplified magnetic beads, Tregs could effectively amplify. Conclusion Immunomagnetic beads can effectively enrich Tregs in umbilical cord blood mononuclear cells without obvious damage. Experiments show that immunomagnetic beads can effectively amplify Tregs in vitro, while evaluation of cells biological function needs further research.

Key words: Regulatory T cells, Immunoamplified bead

中图分类号:

R392.1

苏木雅, 杜华, 师迎旭. 免疫磁珠法体外促脐带血调节性T细胞增殖能力的研究^*[J]. 临床输血与检验, 2020, 22(5): 526-530.

SU Mu-ya, DU Hua, SHI Ying-xu. In Vitro Expansion of Human Umbilical Cord Blood Regulatory T Cells by Magnetic Beads[J]. JOURNAL OF CLINICAL TRANSFUSION AND LABORATORY MEDICINE, 2020, 22(5): 526-530.

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免疫磁珠法体外促脐带血调节性T细胞增殖能力的研究^*

In Vitro Expansion of Human Umbilical Cord Blood Regulatory T Cells by Magnetic Beads

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