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对从新鲜采集的血液与冷藏24小时的血液中分离出的鸡外周血单个核细胞的评估。

Assessment of chicken peripheral blood mononuclear cells isolated from freshly drawn blood versus 24 h refrigerated blood.

作者信息

Tantikositruj Chananphat, Buadkhunthod Anchalee, Rattanasrisomporn Jatuporn, Kitpipit Warangkana, 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. 2021 Sep;14(9):2549-2553. doi: 10.14202/vetworld.2021.2549-2553. Epub 2021 Sep 27.

Abstract

BACKGROUND AND AIM

The peripheral blood mononuclear cell (PBMC) is an excellent cell source for studies, particularly those involving immunology. The aim of this study was to determine the quality and quantity of chicken PBMCs isolated from freshly drawn blood as well as blood that had been chilled for 24 h. In addition, the survival of PBMCs cultured in medium was investigated.

MATERIALS AND METHODS

Blood samples were collected from 12 Betong and 12 Leghorn chickens. Hemograms were analyzed. Density gradient centrifugation was used to isolate PBMCs. PBMCs (2×10 cells/mL) were cultured in a culture medium and incubated in a CO incubator for 5 consecutive days. The number of viable cells was determined using the trypan blue dye exclusion method.

RESULTS

Blood samples were obtained from healthy chickens. There was no statistically significant difference in the total amount of PBMC between fresh and refrigerated blood samples from both chicken breeds. The viability of PBMCs isolated from fresh blood (95%) was significantly greater than blood refrigerated for 24 h (90-92%) in both breeds. Furthermore, the viability of PBMCs isolated from both blood samples decreased significantly over time, from 90-95% to 60-65%.

CONCLUSION

The total number of PBMC in fresh and refrigerated blood was not significantly different. Fresh blood-derived PBMCs had significantly higher viability than 24 h refrigerated blood PBMCs. Furthermore, the viability of PBMCs decreased significantly over time.

摘要

背景与目的

外周血单个核细胞(PBMC)是研究的优良细胞来源,尤其是涉及免疫学的研究。本研究的目的是确定从新鲜采集的血液以及冷藏24小时的血液中分离出的鸡PBMC的质量和数量。此外,还研究了在培养基中培养的PBMC的存活率。

材料与方法

从12只北洞鸡和12只来亨鸡采集血样。分析血常规。采用密度梯度离心法分离PBMC。将PBMC(2×10个细胞/毫升)接种于培养基中,在CO培养箱中连续培养5天。使用台盼蓝染料排斥法测定活细胞数量。

结果

血样取自健康鸡。两个品种的新鲜血样和冷藏血样中PBMC的总量在统计学上无显著差异。两个品种中,从新鲜血液中分离出的PBMC的存活率(95%)显著高于冷藏24小时的血液(90 - 92%)。此外,从两种血样中分离出的PBMC的存活率随时间显著下降,从90 - 95%降至60 - 65%。

结论

新鲜血液和冷藏血液中PBMC的总数无显著差异。新鲜血液来源的PBMC的存活率显著高于冷藏24小时的血液来源的PBMC。此外,PBMC的存活率随时间显著下降。

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本文引用的文献

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Transcriptomes of whole blood and PBMC in chickens.鸡全血和外周血单个核细胞的转录组
Comp Biochem Physiol Part D Genomics Proteomics. 2016 Dec;20:1-9. doi: 10.1016/j.cbd.2016.06.008. Epub 2016 Jun 27.

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