如何高效率转染免疫T细胞？

高效率转染免疫T细胞是大家关心的问题；

一、2020-2-19浙江大学附属第二医院乳腺外科,浙江大学附属第二医院浙江省疗法肿瘤微环境与免疫重点实验室；联合发表标题为Breast cancer-derived exosomes transmit lncRNA SNHG16 to induce CD73+γδ1 Treg cells（乳腺癌衍生的外泌体传递lncRNA SNHG16诱导CD73 +γδ1 Treg细胞）的文章到nature.com/sigtrans杂志，文章已被接受；

二、本研究使用中使用Invigentech（英克）公司INVI DNA RNA 转染试剂转染质粒DNA、siRNA到Vδ1 T?免疫T细胞里面；

三、本文中转染质粒DNA、siRNA，转染细胞数量信息如下：

A.?将全长2435 bp的序列克隆到pCR3.1载体中构建SNHG16过表达载体;

B.?siRNA序列：SNHG16-homo-349，5′GCCUCUGCUGCUAAUUGUUTT-3'; SNHG16-homo-867，?5′-CCAAGGAGGGACUGUUUAATT-3′和SNHG16-homo-2004，5′-CCCAGUGUUGACUCACCAATT-3；

C.?转染细胞用量：6孔板里面1 × 106 cells/well；

四、发表文章部分内容如下：

Breast cancer-derived exosomes transmit lncRNA SNHG16?to induce CD73+γδ1 Treg cells

Plasmid construction and siRNA silencing?A full-length 2435 bp sequence was cloned into the pCR3.1 vector to

construct the SNHG16 overexpression vector. The sequences of?SNHG16-specifific siRNAs were as follows: SNHG16-homo-349, 5′GCCUCUGCUGCUAAUUGUUTT-3′; SNHG16-homo-867, 5′-CCAAGGAGGGACUGUUUAATT-3′ and SNHG16-homo-2004, 5′-CCCAGUGUUGACUCACCAATT-3′. The miR-16–5p mimics, inhibitor and negative

controls were purchased from GenePharma (SupplementaryTable S3).

To manipulate the expression of miR-16–5p in?Vδ1 T cells,?Vδ1T cells were sorted from peripheral blood and seeded into six-wellplates at 1 × 106 cells/well with 1 ml of medium supplementedwith 10% FBS, 10 μg/ml CD3, 10 μg/ml CD28 and 40 U/mL IL-2.Then,?transfection reagent (INVI DNA RNA Transfection Reagent,Invigentech, USA) and miR-16–5p mimics/NC/inhibitor/inhibitor?NC (20 μm) were incubated with the cells for 48 h, after which the?cells were collected for further experiments.

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黄壳HX5 Plus和HX5区别

1.机油油品

黄壳HX5是属于矿物机油，而黄壳HX5 Plus则是半合成机油。不管是机油的使用寿命，还是机油的各项性能，黄壳HX5 Plus都是比黄壳HX5略胜一筹的。

2.机油寿命

黄壳HX5，一般半年或行驶5000公里左右更换一次。而黄壳HX5 Plus，可以每7500公里或者半年更换。但只是参考数值，具体时间还是需要根据机油颜色或者车况进行判断。