A novel Eriocheir sinensis primary hemocyte culture technique and its immunoreactivity after pathogen stimulation
A novel Eriocheir sinensis primary hemocyte culture technique and its immunoreactivity after pathogen stimulation
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中华绒螯蟹原代血细胞培养新技术及其病原体刺激后的免疫反应
DOI：
10.1016/j.aquaculture.2015.04.032
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发表时间：
2015-09
期刊：
影响因子：
4.5
通讯作者：
Qun Wang
中科院分区：
文献类型：
--
作者：
Xing Zhang;Dan Li;Weiwei Li;Qun Wang
为了进一步推进免疫学研究，建立了中华绒螯蟹血细胞的改良原代细胞培养方法，采用不含FBS和co2的L-15培养基，在28°C下培养。在此条件下，培养的细胞稳定繁殖30天。为了评估细胞活性，我们分析了某些关键免疫相关基因的表达水平、血细胞的黑色素化以及细胞信号通路中关键分子的磷酸化状态。LPS、PGN和GLU刺激后，EsDWD1、EsLecG和EsToll2 mRNA表达水平上调。同时，副溶血弧菌的刺激导致血细胞的黑化，并且随着细菌浓度的增加，黑化更加明显。此外，暴露于v。副溶血剂显著诱导JNK、ERK和p38磷酸化，诱导后30min磷酸化程度最大。总的来说，我们的结果建立了一个技术forE。白细胞原代培养并表现出适当的免疫反应、调节和活性。我们声明我们没有利益冲突声明。
In order to advance immunological research, a modified primary cell culture method for hemocytes fromEriocheir sinensis(Chinese mitten crab) was established using L-15 medium lacking FBS and CO2at 28 °C. Under these conditions, cultured cells were stably propagated for 30 days. To evaluate cellular activity, expression levels of certain critical immune-related genes, melanization of the hemocytes, and the phosphorylation status of key molecules in cell signaling pathways were analyzed. mRNA expression level of EsDWD1, EsLecG and EsToll2 was upregulated after LPS, PGN and GLU stimulation. Meanwhile, stimulated byVibrio parahemolyticusled to the melanization of the hemocytes, and the melanization was more obviously with the increasing concentration of bacteria. Moreover, exposure toV. parahemolyticussignificantly induced JNK, ERK and p38 phosphorylation, with the greatest extent of phosphorylation observed 30 min post-induction. Collectively, our results establish a technique forE. sinensishemocyte primary culture and demonstrate appropriate immunological response, regulation and activity.Statement of relevanceWe declare that we have no conflict of interest statement.
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DOI：
10.1007/s11626-010-9343-x
发表时间：
2010-09
期刊：
In Vitro Cellular & Developmental Biology - Animal
影响因子：
--
作者：
S. George;A. Dhar
通讯作者：
S. George;A. Dhar
DOI：
10.1142/9789812707444_0018
发表时间：
2007-02
期刊：
--
影响因子：
--
作者：
S. Bao;Biwen Wu;Hui-qin Huang
通讯作者：
S. Bao;Biwen Wu;Hui-qin Huang
DOI：
10.1023/a:1017971618398
发表时间：
2000
期刊：
Methods in cell science : an official journal of the Society for In Vitro Biology
影响因子：
--
作者：
A. Mulford;F. Lyng;C. Mothersill;B. Austin
通讯作者：
A. Mulford;F. Lyng;C. Mothersill;B. Austin
影响因子：
2.9
作者：
Huang, Xian-De;Yin, Zhi-Xin;He, Jian-Guo
通讯作者：
He, Jian-Guo
DOI：
10.1007/978-94-010-0369-8_62
发表时间：
2001
期刊：
--
影响因子：
--
作者：
G. Kumar;I. Singh;R. Philip
通讯作者：
G. Kumar;I. Singh;R. Philip