总览

通过免疫磁珠负选从新鲜或冻存的人外周血单个核细胞 (PBMCs) 或裂解的白细胞单采术样本中分离出无磁珠标记和高纯度的T细胞。EasySep™无柱免疫磁珠分选技术结合单克隆抗体的特异性和无需分选柱的简便性，20多年来被广泛引用于已发表的文献中。

本EasySep™负选方案通过抗体复合物与磁珠标记非目标细胞，表达以下标志物的非目标细胞将被清除：CD14、CD16、CD19、CD36、CD56、CD66b、CD123及GlyA。通过EasySep™磁极将被磁珠标记的细胞与未被标记的目的细胞分离，接着只需将目的细胞倾倒或吸取至一个新的试管中，仅需8分钟即可获得高纯度的T细胞，且可立即用于流式细胞术、细胞培养或DNA/RNA提取等下游应用。

该产品可替代EasySep™人T细胞富集试剂盒 (产品号 #19051) 以进行更快的细胞分选。

如需从白细胞分离单采术中大规模分离人T细胞，请选用大规格（1x10^10细胞）试剂盒（产品号 #100-0695）

深入了解EasySep™免疫磁珠分选技术原理，或探索RoboSep™全自动免疫磁珠细胞分选方案。亦可选择即用型、符合伦理标准的冻存的人外周血pan-T细胞（使用EasySep™人T细胞分选试剂盒分离）。探索更多为您实验流程优化的产品，包括培养基、添加剂、抗体等。

本EasySep™负选方案通过抗体复合物与磁珠标记非目标细胞，表达以下标志物的非目标细胞将被清除：CD14、CD16、CD19、CD36、CD56、CD66b、CD123及GlyA。通过EasySep™磁极将被磁珠标记的细胞与未被标记的目的细胞分离，接着只需将目的细胞倾倒或吸取至一个新的试管中，仅需8分钟即可获得高纯度的T细胞，且可立即用于流式细胞术、细胞培养或DNA/RNA提取等下游应用。

该产品可替代EasySep™人T细胞富集试剂盒 (产品号 #19051) 以进行更快的细胞分选。

如需从白细胞分离单采术中大规模分离人T细胞，请选用大规格（1x10^10细胞）试剂盒（产品号 #100-0695）

深入了解EasySep™免疫磁珠分选技术原理，或探索RoboSep™全自动免疫磁珠细胞分选方案。亦可选择即用型、符合伦理标准的冻存的人外周血pan-T细胞（使用EasySep™人T细胞分选试剂盒分离）。探索更多为您实验流程优化的产品，包括培养基、添加剂、抗体等。

本EasySep™负选方案通过抗体复合物与磁珠标记非目标细胞，表达以下标志物的非目标细胞将被清除：CD14、CD16、CD19、CD36、CD56、CD66b、CD123及GlyA。通过EasySep™磁极将被磁珠标记的细胞与未被标记的目的细胞分离，接着只需将目的细胞倾倒或吸取至一个新的试管中，仅需8分钟即可获得高纯度的T细胞，且可立即用于流式细胞术、细胞培养或DNA/RNA提取等下游应用。

该产品可替代EasySep™人T细胞富集试剂盒 (产品号 #19051) 以进行更快的细胞分选。

如需从白细胞分离单采术中大规模分离人T细胞，请选用大规格（1x10^10细胞）试剂盒（产品号 #100-0695）

深入了解EasySep™免疫磁珠分选技术原理，或探索RoboSep™全自动免疫磁珠细胞分选方案。亦可选择即用型、符合伦理标准的冻存的人外周血pan-T细胞（使用EasySep™人T细胞分选试剂盒分离）。探索更多为您实验流程优化的产品，包括培养基、添加剂、抗体等。

本EasySep™负选方案通过抗体复合物与磁珠标记非目标细胞，表达以下标志物的非目标细胞将被清除：CD14、CD16、CD19、CD36、CD56、CD66b、CD123及GlyA。通过EasySep™磁极将被磁珠标记的细胞与未被标记的目的细胞分离，接着只需将目的细胞倾倒或吸取至一个新的试管中，仅需8分钟即可获得高纯度的T细胞，且可立即用于流式细胞术、细胞培养或DNA/RNA提取等下游应用。

该产品可替代EasySep™人T细胞富集试剂盒 (产品号 #19051) 以进行更快的细胞分选。

如需从白细胞分离单采术中大规模分离人T细胞，请选用大规格（1x10^10细胞）试剂盒（产品号 #100-0695）

深入了解EasySep™免疫磁珠分选技术原理，或探索RoboSep™全自动免疫磁珠细胞分选方案。亦可选择即用型、符合伦理标准的冻存的人外周血pan-T细胞（使用EasySep™人T细胞分选试剂盒分离）。探索更多为您实验流程优化的产品，包括培养基、添加剂、抗体等。

磁体兼容性
• EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • Easy 50 EasySep™ Magnet (Catalog #18002) • EasyPlate™ EasySep™ Magnet (Catalog 18102) • EasyEights™ EasySep™ Magnet (Catalog #18103) • RoboSep™-S (Catalog #21000) • Easy 250 EasySep™ Magnet (Catalog #100-0821)

亚型
细胞分选试剂盒

细胞类型
T 细胞

种属
人

样本来源
Leukapheresis，PBMC

筛选方法
Negative

应用
细胞分选

品牌
EasySep，RoboSep

研究领域
嵌合体，HLA，免疫，细胞治疗开发

查看更多显示更少

实验数据

T Cell Separation using EasySep™ Human T Cell Isolation Kit

Figure 1. EasySep™ Human T Cell Isolation Kit

Starting with human peripheral blood mononuclear cells (PBMCs), the T cell content (CD3+) of the isolated fraction is typically 96.7 ± 1.5% (mean ± SD).

Figure 2. ImmunoCult™-XF T Cell Expansion Medium Supports Faster T Cell Expansion Than Other Serum-Free and Serum-Supplemented Media

T cells were isolated from human peripheral blood samples using the EasySep™ Human T Cell Isolation Kit (Catalog #17951), stimulated with ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator (Catalog #10970), and cultured in ImmunoCult™-XF T Cell Expansion Medium supplemented with rhIL-2. T cells were stimulated with ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator on Day 0 and every 7 to 8 days for the duration of the culture. T cells were analyzed on Days 4, 7, 8, 10, 11, 14, 18, and 21 for fold expansion relative to the initial cell seeding density. Compared to all competitor media tested, ImmunoCult™-XF T Cell Expansion Medium showed significantly higher expansion of total T cells. Commercial alternatives 1 to 4 include, in no particular order, X-VIVO™ 15 (Lonza), AIM V® Medium (Life Tech), CellGro® DC Medium (CellGenix), and RPMI 1640 + serum. Each data point represents the mean fold expansion ± S.E.M. at the specified time points (p<0.05 for ImmunoCult™-XF versus all media for Days 8, 11, 14, 18, and 21, tested using two-tailed, paired t-test with unequal variance, n = 6 to 19 donors). The average fold expansion of T cells in ImmunoCult™-XF T Cell Expansion Medium were 15-fold on Day 7, 80-fold on Day 10, 450-fold on Day 14, and 4,000-fold on Day 21.

Figure 3. T Cell Expansion and Activation Ability Is Better Preserved When Leukopaks are Stored at Fridge Temperature

Using EasySep™ Human T Cell Isolation Kit (Catalog #17951), T cells were isolated from 1 leukopak fraction (Catalog # 70500) of each storage condition daily for 5 days, and 1 x 10⁶ isolated cells were cultured in ImmunoCult™-XF T Cell Expansion Medium (Catalog #10981) supplemented with ImmunoCult™ Human CD3/CD28/CD2 T Cell Activator (Catalog # 10970) and 10 ng/mL IL-2 for 10 days with assessment of CD25 activation marker expression on day 3 of culture. (A) Representative flow cytometry data from leukopaks stored 1 day at fridge temperature (FT), showing that both CD4+ and CD8+ cells are CD25-negative at the start of culture (Day 0), and upregulate CD25 expression by Day 3 of culture. (B) Cellular expansion and corresponding cell yield over 7 days of culture decreases in correlation with storage duration of leukopak fractions. Leukopaks that were stored for 1 - 2 days at either room temperature (RT) or FT had high expansion potential yielding 1.5 - 2 x 10⁷ cells, and this 15 - 20-fold expansion potential is maintained in T cells from leukopaks stored at FT for up to 5 days. In contrast, little or no expansion is observed in T cell cultures from leukopaks stored at RT for 3 or more days, indicating a loss of proliferative capacity. Moreover, T cells show a gradual reduction in their ability to become activated by ImmunoCult™ T Cell activator, as shown by a reduction in Day 3 CD25 expression in gated CD4+ (bottom left) or CD8+ (bottom right) cells over time, and the effect is most pronounced with storage at RT for 3 or more days. All data points represent average ± standard deviation values from leukopak fractions of n = 3 unique donors.

ImmunoCult™ Human T Cell Activators Can Be Used to Optimize Culture Conditions for High-Efficiency TRAC Knockout from Human Primary T Cells

Figure 4. ImmunoCult™ Human T Cell Activators Can Be Used to Optimize Culture Conditions for High-Efficiency TRAC Knockout from Human Primary T Cells

The TRAC locus of human primary T cells was edited with an RNP-based-CRISPR-Cas9 system using multiple T cell activation reagents and dynamics then evaluated to identify a condition with the highest knockout efficiency. (A) TRAC knockout efficiency in human T cells isolated using EasySep™ Human T Cell Isolation Kit (Catalog #17951) and activated with either ImmunoCult™ Human CD3/CD28 or CD3/CD28/CD2 T Cell Activator (Catalog #10971/10970) for 2 or 3 days was assessed by binding the TCRαβ and CD3 receptors with antibodies and performing flow cytometry analysis. Each data point per condition represents an individual donor; n = 4 - 8 donors. Error bars represent standard error of the mean. (B) Genome editing (cleavage) efficiency was assessed at 48 hours post electroporation in human T cells activated with ImmunoCult™ Human CD3/CD28 T Cell Activator for 3 days using the ArciTect™ T7 Endonuclease I Kit (Catalog #76021). Mock electroporated: - RNP; RNP electroporated: + RNP. (C - D) Representative dot plots of TCRαβ and CD3 flow cytometry analysis from (C) mock electroporated and (D) RNP electroporated human T cells activated with ImmunoCult™ Human CD3/CD28 T Cell Activator for 3 days. (E) Representative dot plot of CD4 and CD8 flow cytometry analysis of human T cells activated with ImmunoCult™ Human CD3/CD28 T Cell Activator for 3 days.

产品说明书及文档

请在《产品说明书》中查找相关支持信息和使用说明，或浏览下方更多实验方案。

Document Type

Product Name

Catalog #

Lot #

Language

Document Type

Product Information Sheet

Product Name

RoboSep™ Human T Cell Isolation Kit

Catalog #

17951RF

Lot #

All

Language

English

Document Type

Product Information Sheet

Product Name

EasySep™ Human T Cell Isolation Kit

Catalog #

17951

Lot #

All

Language

English

Document Type

Product Information Sheet

Product Name

EasySep™ Human T Cell Isolation Kit

Catalog #

100-0695

Lot #

All

Language

English

Document Type

Safety Data Sheet 1

Product Name

RoboSep™ Human T Cell Isolation Kit

Catalog #

17951RF

Lot #

All

Language

English

Document Type

Safety Data Sheet 2

Product Name

RoboSep™ Human T Cell Isolation Kit

Catalog #

17951RF

Lot #

All

Language

English

Document Type

Safety Data Sheet 3

Product Name

RoboSep™ Human T Cell Isolation Kit

Catalog #

17951RF

Lot #

All

Language

English

Document Type

Safety Data Sheet 1

Product Name

EasySep™ Human T Cell Isolation Kit

Catalog #

17951

Lot #

All

Language

English

Document Type

Safety Data Sheet 2

Product Name

EasySep™ Human T Cell Isolation Kit

Catalog #

17951

Lot #

All

Language

English

Document Type

Safety Data Sheet 1

Product Name

EasySep™ Human T Cell Isolation Kit

Catalog #

100-0695

Lot #

All

Language

English

Document Type

Safety Data Sheet 2

Product Name

EasySep™ Human T Cell Isolation Kit

Catalog #

100-0695

Lot #

All

Language

English

应用领域

本产品专为以下研究领域设计，适用于工作流程中的高亮阶段。探索这些工作流程，了解更多我们为各研究领域提供的其他配套产品。

Research Area

Workflow Stages

Workflow Stages for T Cell Research

Cell Sourcing

Cell Isolation

Cell Activation, Expansion, and Differentiation

Cell Characterization

Workflow Stages for T Cell Engineering

Cell Isolation

Cell Activation, Expansion, and Differentiation

Cell Characterization

Cell Cryopreservation

Workflow Stages for T Cell Therapy Development

View All

Can EasySep™ be used for either positive or negative selection?

Yes. The EasySep™ kits use either a negative selection approach by targeting and removing unwanted cells or a positive selection approach targeting desired cells. Depletion kits are also available for the removal of cells with a specific undesired marker (e.g. GlyA).

How does the separation work?

Magnetic particles are crosslinked to cells using Tetrameric Antibody Complexes (TAC). When placed in the EasySep™ Magnet, labeled cells migrate to the wall of the tube. The unlabeled cells are then poured off into a separate fraction.

Which columns do I use?

The EasySep™ procedure is column-free. That's right - no columns!

How can I analyze the purity of my enriched sample?

The Product Information Sheet provided with each EasySep™ kit contains detailed staining information.

Can EasySep™ separations be automated?

Yes. RoboSep™, the fully automated cell separator, automates all EasySep™ labeling and cell separation steps.

Can EasySep™ be used to isolate rare cells?

Yes. We recommend a cell concentration of 2x10⁸ cells/mL and a minimum working volume of 100 µL. Samples containing 2x10⁷ cells or fewer should be suspended in 100 µL of buffer.

Are the EasySep™ magnetic particles FACS-compatible?

Yes, the EasySep™ particles are flow cytometry-compatible, as they are very uniform in size and about 5000X smaller than other commercially available magnetic beads used with column-free systems.

Can the EasySep™ magnetic particles be removed after enrichment?

No, but due to the small size of these particles, they will not interfere with downstream applications.

Can I alter the separation time in the magnet?

Yes; however, this may impact the kit's performance. The provided EasySep™ protocols have already been optimized to balance purity, recovery and time spent on the isolation.

For positive selection, can I perform more than 3 separations to increase purity?

Yes, the purity of targeted cells will increase with additional rounds of separations; however, cell recovery will decrease.

How does the binding of the EasySep™ magnetic particle affect the cells? is the function of positively selected cells altered by the bound particles?

Hundreds of publications have used cells selected with EasySep™ positive selection kits for functional studies. Our in-house experiments also confirm that selected cells are not functionally altered by the EasySep™ magnetic particles.

If particle binding is a key concern, we offer two options for negative selection. The EasySep™ negative selection kits can isolate untouched cells with comparable purities, while RosetteSep™ can isolate untouched cells directly from whole blood without using particles or magnets.

IL-6 drives T cell death to participate in lymphopenia in COVID-19. X. Zhou et al. International immunopharmacology 2022 oct

Abstract

Lymphopenia is a common observation in patients with COVID-19. To explore the cause of T cell lymphopenia in the disease, laboratory results of 64 hospitalized COVID-19 patients were retrospectively analyzed and six patients were randomly selected to trace their changes of T lymphocytes and plasma concentration of IL-6 for the course of disease. Results confirmed that the T-cell lymphopenia, especially CD4+ T cell reduction in COVID-19 patients, was a reliable indicator of severity and hospitalization in infected patients. And CD4+ T cell count below 200 cells/$\mu$L predicts critical illness in COVID-19 patients. In vitro assay supported that exposure to key contributors (IL-1$\beta$, IL-6, TNF-$\alpha$ and IFN-$\gamma$) of COVID-19 cytokine storm caused substantial death of activated T cells. Among these contributors, IL-6 level was found to probably reversely correlate with T cell counts in patients. And IL-6 alone was potent to induce T cell reduction by gasderminE-mediated pyroptosis, inferring IL-6 took a part in affecting the function and status of T cells in COVID-19 patients. Intervention of IL-6 mediated T cell pryprotosis may effectively delay disease progression, maintain normal immune status at an early stage of infection.

View publication

Expanding anti-CD38 immunotherapy for lymphoid malignancies. X. Wang et al. Journal of experimental & clinical cancer research : CR 2022 jun

Abstract

BACKGROUND Lymphoid neoplasms, including multiple myeloma (MM), non-Hodgkin lymphoma (NHL), and NK/T cell neoplasms, are a major cause of blood cancer morbidity and mortality. CD38 (cyclic ADP ribose hydrolase) is a transmembrane glycoprotein expressed on the surface of plasma cells and MM cells. The high expression of CD38 across MM and other lymphoid malignancies and its restricted expression in normal tissues make CD38 an attractive target for immunotherapy. CD38-targeting antibodies, like daratumumab, have been approved for the treatment of MM and tested against lymphoma and leukemia in multiple clinical trials. METHODS We generated chimeric antigen receptor (CAR) T cells targeting CD38 and tested its cytotoxicity against multiple CD38high and CD38low lymphoid cancer cells. We evaluated the synergistic effects of all-trans retinoic acid (ATRA) and CAR T cells or daratumumab against cancer cells and xenograft tumors. RESULTS CD38-CAR T cells dramatically inhibited the growth of CD38high MM, mantle cell lymphoma (MCL), Waldenstrom's macroglobulinemia (WM), T-cell acute lymphoblastic leukemia (T-ALL), and NK/T-cell lymphoma (NKTCL) in vitro and in mouse xenografts. ATRA elevated CD38 expression in multiple CD38low cancer cells and enhanced the anti-tumor activity of daratumumab and CD38-CAR T cells in xenograft tumors. CONCLUSIONS These findings may expand anti-CD38 immunotherapy to a broad spectrum of lymphoid malignancies and call for the incorporation of ATRA into daratumumab or other anti-CD38 immunological agents for cancer therapy.

View publication

Indomethacin-induced oxidative stress enhances death receptor 5 signaling and sensitizes tumor cells to adoptive T-cell therapy. N. S. Aboelella et al. Journal for immunotherapy of cancer 2022 jul

Abstract

BACKGROUND Adoptive cell therapy (ACT) using genetically modified T cells has evolved into a promising treatment option for patients with cancer. However, even for the best-studied and clinically validated CD19-targeted chimeric antigen receptor (CAR) T-cell therapy, many patients face the challenge of lack of response or occurrence of relapse. There is increasing need to improve the efficacy of ACT so that durable, curative outcomes can be achieved in a broad patient population. METHODS Here, we investigated the impact of indomethacin (indo), a non-steroidal anti-inflammatory drug (NSAID), on the efficacy of ACT in multiple preclinical models. Mice with established B-cell lymphoma received various combinations of preconditioning chemotherapy, infusion of suboptimal dose of tumor-reactive T cells, and indo administration. Donor T cells used in the ACT models included CD4+ T cells expressing a tumor-specific T cell receptor (TCR) and T cells engineered to express CD19CAR. Mice were monitored for tumor growth and survival. The effects of indo on donor T cell phenotype and function were evaluated. The molecular mechanisms by which indo may influence the outcome of ACT were investigated. RESULTS ACT coupled with indo administration led to improved tumor growth control and prolonged mouse survival. Indo did not affect the activation status and tumor infiltration of the donor T cells. Moreover, the beneficial effect of indo in ACT did not rely on its inhibitory effect on the immunosuppressive cyclooxygenase 2 (COX2)/prostaglandin E2 (PGE2) axis. Instead, indo-induced oxidative stress boosted the expression of death receptor 5 (DR5) in tumor cells, rendering them susceptible to donor T cells expressing TNF-related apoptosis-inducing ligand (TRAIL). Furthermore, the ACT-potentiating effect of indo was diminished against DR5-deficient tumors, but was amplified by donor T cells engineered to overexpress TRAIL. CONCLUSION Our results demonstrate that the pro-oxidative property of indo can be exploited to enhance death receptor signaling in cancer cells, providing rationale for combining indo with genetically modified T cells to intensify tumor cell killing through the TRAIL-DR5 axis. These findings implicate indo administration, and potentially similar use of other NSAIDs, as a readily applicable and cost-effective approach to augment the efficacy of ACT.

View publication

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EasySep™人T细胞分选试剂盒

产品号 #（选择产品）

产品号 #17951_C

产品优势

产品组分包括

What Our Scientist Says

总览

实验数据

产品说明书及文档

应用领域

相关材料与文献

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Scientists Helping Scientists™

Molecular Weight	1645,1646
种属	Human
Magnet Compatibility	• EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • Easy 50 EasySep™ Magnet (Catalog #18002) • EasyPlate™ EasySep™ Magnet (Catalog 18102) • EasyEights™ EasySep™ Magnet (Catalog #18103) • RoboSep™-S (Catalog #21000) • Eas
样本来源	Leukapheresis, PBMC
Selection Method	Negative
Target	• EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • Easy 50 EasySep™ Magnet (Catalog #18002) • EasyPlate™ EasySep™ Magnet (Catalog 18102) • EasyEights™ EasySep™ Magnet (Catalog #18103) • RoboSep™-S (Catalog #21000) • Eas
Pathway	Negative

EasySep™人T细胞分选试剂盒

产品号 #（选择产品）

产品号 #17951_C

产品优势

产品组分包括

What Our Scientist Says

总览

实验数据

产品说明书及文档

应用领域

相关材料与文献

技术资料 (26)

常见问题

Can EasySep™ be used for either positive or negative selection?

How does the separation work?

Which columns do I use?

How can I analyze the purity of my enriched sample?

Can EasySep™ separations be automated?

Can EasySep™ be used to isolate rare cells?

Are the EasySep™ magnetic particles FACS-compatible?

Can the EasySep™ magnetic particles be removed after enrichment?

Can I alter the separation time in the magnet?

For positive selection, can I perform more than 3 separations to increase purity?

How does the binding of the EasySep™ magnetic particle affect the cells? is the function of positively selected cells altered by the bound particles?

文献 (20)

Abstract

Abstract

Abstract

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