StemCell 19051 EasySep™ Enrichment Kit 人T细胞富...

推荐使用新版EasySep™人T细胞分选试剂盒（货号：17951），短至8分钟即可完成免疫磁珠细胞分选。

CAR-T是一种通过基因工程改造T细胞对白血病的血液肿瘤细胞进行攻击的免疫疗法。截止今年十月，已有诺华和Kite两家公司获得FDA认证的唯二基因疗法。CAR-T免疫细胞疗法大热中。这两个临床成功的CAR-T疗法都对来自病人的外周血T细胞进行分选。

Stemcell的EasySep™人T细胞富集试剂盒可以用于CAR-T疗法的T细胞分选阶段（免疫磁珠细胞分选法）^2，3。EasySep™人T细胞富集试剂盒的磁珠分选采用负选方式，可以避免标记T细胞，以防标记抗体对T细胞造成伤害，Z大程度保持T细胞的untouched活性，立即用于下游CAR-T研究。

基本信息

中文名称 EasySep™人T细胞富集试剂盒

英文名称 EasySep™ Human T Cell Enrichment Kit

用于标记1 x 10⁹个细胞

EasySep™人T细胞富集试剂盒是套装，包含：

EasySep™ Human T Cell Enrichment Cocktail（1mL） T细胞富集抗体复合物

EasySep™ D Magnetic Particles（1mL） 磁珠

货号：19051

保存 2 - 8°C，不能冷冻。

产品订购信息

说明

RF：货号后加RF，比如17951RF，代表RoboSep™试剂盒：包含EasySep™分选试剂盒、RoboSep™缓冲液和1-2盒RoboSep™枪头。

分选试剂盒和富集试剂盒的主要区别：分选试剂盒是富集试剂盒的更新版，实验耗时更少。

>>>>>>>>>>>>>>>>>>>>>>>>>订购直达的分割线>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>

选购19051 Stemcell EasySep™ Enrichment Kit CAR-T疗法免疫细胞磁珠富集分选试剂盒欢迎您致电 华雅再生医学旗舰公司：红荣微再（上海）生物工程技术有限公司 ：152 1681 4001。华雅再生医学-客服: 316 808 6348。

EasySep™人T细胞富集试剂盒应用CAR-T免疫细胞疗法流程的相关阶段

病人来源的T细胞样本（PBMC等）——T细胞分离（免疫磁珠细胞分选法，EasySep™人T细胞富集试剂盒）——CAR-T细胞活化、扩增和基因编辑——CAR-T细胞回输

Stemcell EasySep™人T细胞富集试剂盒适用样本

EasySep™人T细胞富集试剂盒可用于从各种样本来源中分选高纯度的人T细胞，包括：新鲜或冻存外周血单个核细胞（PBMCs）、白细胞单采术（Leuko Pak）样本。目标细胞被抗体复合物和磁珠标记，以进行负选。

诺华的CAR-T免疫细胞疗法使用同样的样本¹。

Stemcell EasySep™人T细胞富集试剂盒优点

方便快捷，新版短至8分钟。无柱分选

纯度Z高达99%

细胞保持Untouched

Stemcell EasySep Human T Cell Enrichment Kit组份

PBS - phosphate-buffered saline; TBS - TRIS-buffered saline

Components may be shipped at room temperature (15 - 25°C) but should be stored as indicated above.

Stemcell EasySep™人T细胞富集试剂盒的免疫磁珠细胞分选常规操作流程（负选）

1.将EasySep™分选抗体混合物加入到单细胞悬液中，并孵育3分钟

2.将EasySep™ 磁珠加入到细胞悬液中。

3.将试管放置在EasySep™磁极（货号18000）中，并孵育3分钟。

4.将目标细胞倾倒至新的试管中。

*该时间是新一代的负选试剂盒所需的常规时间。每款试剂盒所需的实际时间取决于使用的磁极和分选流程。新一代的负选流程中无需磁珠孵育的步骤。

T细胞负选方案介绍

在负选时，非目标细胞通过针对细胞表面抗原的抗体被去除。负选试剂中的抗体组合确定哪些细胞被去除。

以下情况，比较适合负选：

需要未经标记的细胞

若将抗体结合到目标细胞上会造成YZ或活化，影响下游研究

目标细胞没有特异性抗体可用来标记

需要去除某些类型的细胞

文献——使用Stemcell EasySep™人T细胞富集试剂盒进行CAR-T疗法的免疫磁珠细胞分选

Stemcell EasySep Human T Cell Enrichment Kit免疫磁珠细胞分选相关产品

概要

The EasySep™ Human T Cell Enrichment Kit is designed to isolate T cells from fresh or previously frozen peripheral blood mononuclear cells by negative selection. Unwanted cells are targeted for removal with Tetrameric Antibody Complexes recognizing non-T cells and dextran-coated magnetic particles. The labeled cells are separated using an EasySep™ magnet without the use of columns. Desired cells are poured off into a new tube.

For even faster cell isolations, we recommend the new EasySep™ Human T Cell Isolation Kit (17951) which isolates cells in just 8 minutes.

Stemcell EasySep™人T细胞富集试剂盒说明书

Description

Isolate untouched and highly purified T cells from fresh or previously frozen human peripheral blood mononuclear cells (PBMCs) or lysed leukapheresis samples

by immunomagnetic negative selection.

– Fast, easy-to-use and column-free

– Up to 99% purity

– Untouched, viable cells

EasySep™ Human T Cell Enrichment Kit targets non-T cells for removal with antibodies recognizing specific cell surface markers. Unwanted cells are labeled with antibodies and magnetic particles, and separated without columns using an EasySep™ magnet. Desired cells are simply poured off into a new tube. Isolated cells are immediay available for downstream applications such as flow cytometry, culture, or DNA/RNA extraction.

EasySep™ Human T Cell Enrichment Kit-样本制备Sample Preparation

PERIPHERAL BLOOD

Prepare a PBMC suspension from whole peripheral blood by centrifugation over a density gradient medium (e.g. Lymphoprep™, Catalog #07801). For more rapid PBMC preparation, use the SepMate™ RUO (Catalog #86450/86415) or SepMate™ IVD* (Catalog #85450/85415) cell isolation tube.

If using previously frozen PBMCs, incubate the cells with DNase I Solution (Catalog #07900) at a concentration of 100 μg/mL at room temperature (15 - 25°C) for at least 15 minutes prior to labeling and separation. Filter aggregated suspensions through a 40 μm Cell Strainer (Catalog #27305) for optimal results.

After preparation, resuspend cells at 5 x 10^7 cells/mL in recommended medium.

* SepMate™ IVD is only available in select regions where it is registered as an In Vitro Diagnostic (IVD) device for the isolation of mononuclear cells (MNCs) from whole blood or bone marrow by density gradient centrifugation. In all other regions SepMate™ is available for research use only (RUO).

白细胞去除术LEUKAPHERESIS (LEUKO PAK)

If working with large volumes (> 150 mL), concentrate leukapheresis cells first by centrifuging at 500 x g for 10 minutes. Remove the supernatant and resuspend the cells in 1/10th of the original leukapheresis volume with recommended medium (e.g. for 300 mL of cells, resuspend in 30 mL of recommended medium). For small volumes (≤ 150 mL), add Ammonium Chloride Solution (Catalog #07800) directly to the leukapheresis sample.

1. Add an equal volume of Ammonium Chloride Solution to the leukapheresis sample.

2. Incubate on ice for 15 minutes.

3. Centrifuge at 500 x g for 10 minutes at room temperature (15 - 25°C). Remove the supernatant.

4. Wash the cells by topping up the tube with recommended medium. Centrifuge the cells at 150 x g for 10 minutes at room temperature with the brake off.

Carefully remove the supernatant.

5. Repeat step 4 one or more times until most of the plaets have been removed (indicated by a clear supernatant).

6. Resuspend the cells at 5 x 10^7 cells/mL in recommended medium.

EasySep™ Human T Cell Enrichment Kit推荐介质Recommended Medium

EasySep™ Buffer (Catalog #20144), RoboSep™ Buffer (Catalog #20104), or PBS containing 2% fetal bovine serum (FBS) and 1 mM EDTA. Medium should be free of Ca++ and Mg++.

CAR-T免疫细胞疗法参考资料：

1. (2017) Global Manufacturing of CAR T Cell Therapy.Bruce L.Levine et al.Volume 4, 17 March 2017, Pages 92-101.

2. (2017) Targeting alpha-fetoprotein(AFP)-MHC complex with CAR T-cell therapy for liver cancer. Liu H et al.Clin Cancer Res 23(2): 478-88.

3. (2014) An NCR1-based chimeric receptor endows T-cells with multiple anti-tumor specificities. Tal Y et al.Oncotarget 5(21): 10949-58.

19051 Stemcell EasySep™ Enrichment Kit CAR-T疗法免疫细胞磁珠富集分选试剂盒产品资料由红荣微再编辑整理。红荣微再（上海）生物工程技术有限公司以“传递科学价值，服务科学研究”为宗旨，主营：干细胞、JZYL、细胞ZL、器官再生 四大板块的产品。

选购19051 Stemcell EasySep™ Enrichment Kit CAR-T疗法免疫细胞磁珠富集分选试剂盒欢迎您致电 华雅再生医学旗舰公司：红荣微再（上海）生物工程技术有限公司 ：152 1681 4001。华雅再生医学-客服: 316 808 6348。