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北洞鸡外周血单个核细胞的分离及Toll样受体的表达

Isolation of peripheral blood mononuclear cells and the expression of toll-like receptors in Betong chickens.

作者信息

Suklek Anutian, Kayan Autchara, Rattanasrisomporn Jatuporn, Boonkaewwan Chaiwat

机构信息

Department of Animal Science, Faculty of Agriculture, Kasetsart University, Bangkok 10900, Thailand.

Department of Companion Animal Clinical Science, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand.

出版信息

Vet World. 2020 Jul;13(7):1372-1375. doi: 10.14202/vetworld.2020.1372-1375. Epub 2020 Jul 18.

Abstract

BACKGROUND AND AIM

Toll-like receptors (TLRs) comprise microbial sensing receptors present on cell surfaces that are capable of detecting pathogens. The present study aims to examine the expression of TLRs within the peripheral blood mononuclear cell (PBMC) of the Betong chickens.

MATERIALS AND METHODS

Blood samples were harvested from 12 Betong (KU line) chickens. Hematological values were calculated. PBMC was isolated from the blood utilizing a Histopaque solution and stored in a RPMI1640 culture medium. Cell viability was investigated using a Trypan Blue dye exclusion test. DNA was extracted from PBMC and the expression of the DNA's TLRs was examined using a polymerase chain reaction.

RESULTS

Hematological values were determined from the blood samples collected in this study obtained from healthy Betong chickens. PBMC that was isolated from the Betong chickens possessed cell viability higher than 95% (95.37±1.06). From the examination of TLRs gene expression, results revealed instances of TLR1.1, TLR1.2, TLR2.1, TLR2.2, TLR3, TLR4, TLR5, TLR 7, TLR15, and TLR21 that were present in the PBMC of Betong chickens.

CONCLUSION

PBMC isolated from the blood of healthy Betong chickens possessed excellent cell quality. All chicken TLRs were discovered within the PBMC of Betong chickens. Hence, PBMC stands out as one of the premier sources for studies of chicken immune response.

摘要

背景与目的

Toll样受体(TLRs)是存在于细胞表面的微生物传感受体,能够检测病原体。本研究旨在检测北洞鸡外周血单个核细胞(PBMC)中TLRs的表达。

材料与方法

采集12只北洞(KU系)鸡的血液样本。计算血液学指标。利用Histopaque溶液从血液中分离PBMC,并保存在RPMI1640培养基中。使用台盼蓝染料排除试验研究细胞活力。从PBMC中提取DNA,并使用聚合酶链反应检测DNA中TLRs的表达。

结果

从本研究收集的健康北洞鸡血液样本中测定血液学指标。从北洞鸡分离的PBMC细胞活力高于95%(95.37±1.06)。通过检测TLRs基因表达,结果显示北洞鸡PBMC中存在TLR1.1、TLR1.2、TLR2.1、TLR2.2、TLR3、TLR4、TLR5、TLR 7、TLR15和TLR21。

结论

从健康北洞鸡血液中分离的PBMC具有优良的细胞质量。在北洞鸡的PBMC中发现了所有鸡TLRs。因此,PBMC是研究鸡免疫反应的主要来源之一。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9676/7429389/da729ce8b1db/Vetworld-13-1372-g001.jpg

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