EasySep™人T細胞富集試劑盒可用于從新鮮或凍存外周血單個核細胞中通過負選方式分離T細胞。不需要的細胞被識別非T細胞的四聚體抗體復(fù)合物（Tetrameric Antibody Complexes）和dextran包被的磁珠標(biāo)記后移除。被標(biāo)記細胞不需要過柱，使用EasySep™磁極就能簡單分離。需要的細胞被倒到新管子。分離細胞可以立即用于下游：流式細胞術(shù)、細胞培養(yǎng)（CAR-T免疫療法）或DNA/RNA提取。
推薦使用新版EasySep™人T細胞分選試劑盒（貨號：17951），短至8分鐘即可完成免疫磁珠細胞分選。
CAR-T是一種通過基因工程改造T細胞對白血病的血液腫瘤細胞進行攻擊的免疫療法。截止今年十月，已有諾華和Kite兩家公司獲得FDA認證的唯二基因療法。CAR-T免疫細胞療法大熱中。這兩個臨床成功的CAR-T療法都對來自病人的外周血T細胞進行分選。Stemcell的EasySep™人T細胞富集試劑盒可以用于CAR-T療法的T細胞分選階段（免疫磁珠細胞分選法）2，3。EasySep™人T細胞富集試劑盒的磁珠分選采用負選方式，可以避免標(biāo)記T細胞，以防標(biāo)記抗體對T細胞造成傷害，zui大程度保持T細胞的untouched活性，立即用于下游CAR-T研究。
基本信息
中文名稱 EasySep™人T細胞富集試劑盒
英文名稱 EasySep™ Human T Cell Enrichment Kit
用于標(biāo)記1 x 109個細胞
EasySep™人T細胞富集試劑盒是套裝，包含：
EasySep™ Human T Cell Enrichment Cocktail（1mL） T細胞富集抗體復(fù)合物
EasySep™ D Magnetic Particles（1mL） 磁珠
貨號：19051
保存 2 - 8°C，不能冷凍。
產(chǎn)品訂購信息
細胞類型 | 貨號 | 產(chǎn)品名稱 | 純度 | 適用的染色抗體 |
T細胞 | 17951 17951RF | EasySep Human T Cell Isolation Kit EasySep™人T細胞分選試劑盒 | 96.7 ± 1.5% | CD2（貨號#60007） CD3（貨號#60011） CD4（貨號#60016） CD8a（貨號#60022） CD45（貨號#60018） |
19051 19051RF | EasySep Human T Cell Enrichment Kit EasySep™人T細胞富集試劑盒 | 95.0 - 99.0% |
說明
RF：貨號后加RF，比如17951RF，代表RoboSep™試劑盒：包含EasySep™分選試劑盒、RoboSep™緩沖液和1-2盒RoboSep™槍頭。
分選試劑盒和富集試劑盒的主要區(qū)別：分選試劑盒是富集試劑盒的更新版，實驗耗時更少。
>>>>>>>>>>>>>>>>>>>>>>>>>訂購直達的分割線>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>
選購19051 Stemcell EasySep™ Enrichment Kit CAR-T療法免疫細胞磁珠富集分選試劑盒歡迎您致電 華雅再生醫(yī)學(xué)旗艦公司：紅榮微再（上海）生物工程技術(shù)有限公司 ：152 1681 4001。華雅再生醫(yī)學(xué)-客服: 316 808 6348。
EasySep™人T細胞富集試劑盒應(yīng)用CAR-T免疫細胞療法流程的相關(guān)階段
病人來源的T細胞樣本（PBMC等）——T細胞分離（免疫磁珠細胞分選法，EasySep™人T細胞富集試劑盒）——CAR-T細胞活化、擴增和基因編輯——CAR-T細胞回輸
Stemcell EasySep™人T細胞富集試劑盒適用樣本
EasySep™人T細胞富集試劑盒可用于從各種樣本來源中分選高純度的人T細胞，包括：新鮮或凍存外周血單個核細胞（PBMCs）、白細胞單采術(shù)（Leuko Pak）樣本。目標(biāo)細胞被抗體復(fù)合物和磁珠標(biāo)記，以進行負選。
諾華的CAR-T免疫細胞療法使用同樣的樣本1。
Stemcell EasySep™人T細胞富集試劑盒優(yōu)點
方便快捷，新版短至8分鐘。無柱分選
純度zui高達99%
細胞保持Untouched
Stemcell EasySep Human T Cell Enrichment Kit組份
成分 | 貨號 | 數(shù)量 | 保存 | 保質(zhì)期 | 內(nèi)容 |
EasySep™ Human T Cell Enrichment Cocktail EasySep™分選抗體混合物 | 19051C.2 | 1 x 1 mL | 保存 2 - 8°C，不能冷凍 | 效期內(nèi)穩(wěn)定 | 單克隆抗體復(fù)合物（加PBS） |
EasySep™ D Magnetic Particles EasySep™ 磁珠 | 19250 | 1 x 1 mL | 保存 2 - 8°C，不能冷凍 | 效期內(nèi)穩(wěn)定 | 磁珠懸液（加TBS） |
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細胞富集試劑盒的免疫磁珠細胞分選常規(guī)操作流程（負選）
1.將EasySep™分選抗體混合物加入到單細胞懸液中，并孵育3分鐘
2.將EasySep™ 磁珠加入到細胞懸液中。
3.將試管放置在EasySep™磁極（貨號18000）中，并孵育3分鐘。
4.將目標(biāo)細胞傾倒至新的試管中。
*該時間是新一代的負選試劑盒所需的常規(guī)時間。每款試劑盒所需的實際時間取決于使用的磁極和分選流程。新一代的負選流程中無需磁珠孵育的步驟。
T細胞負選方案介紹
在負選時，非目標(biāo)細胞通過針對細胞表面抗原的抗體被去除。負選試劑中的抗體組合確定哪些細胞被去除。
以下情況，比較適合負選：
需要未經(jīng)標(biāo)記的細胞
若將抗體結(jié)合到目標(biāo)細胞上會造成抑制或活化，影響下游研究
目標(biāo)細胞沒有特異性抗體可用來標(biāo)記
需要去除某些類型的細胞
文獻——使用Stemcell EasySep™人T細胞富集試劑盒進行CAR-T療法的免疫磁珠細胞分選
文獻 | 疾病 | 靶點 | CAR-T細胞工程 | 細胞分選產(chǎn)品 | 貨號 |
2 | zui常見肝癌HCC（hepatocellular carcinoma ） | HCC特異性分泌蛋白抗原（alpha-fetoprotein ，AFP） | —— | EasySep™ Human T Cell Isolation Kit，19051 EasySep™人T細胞富集試劑盒的新版 | 17951 17951RF |
3 | 多種腫瘤：carcinomas, neuroblastomas and leukemias等 | 多種腫瘤 | 表達自然殺傷細胞受體1（NCR1 ，natural cytotoxic receptor 1） | EasySep™ Human T Cell Isolation Kit， 19051 EasySep™人T細胞富集試劑盒的新版 | 17951 17951RF |
Stemcell EasySep Human T Cell Enrichment Kit免疫磁珠細胞分選相關(guān)產(chǎn)品
品牌 | 貨號 | 產(chǎn)品描述 |
Stemcell | 17951 | EasySep™ Human T Cell Isolation Kit，EasySep™人T細胞富集試劑盒的新版 |
Stemcell | 17851 | EasySep™ Human CD3 Positive Selection Kit II，T細胞正選 |
| ||
Stemcell | 18000 | EasySep™磁極 |
Stemcell | 20144 | EasySep™ Buffer |
Stemcell EasySep Human T Cell Enrichment Kit英文版介紹
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細胞富集試劑盒說明書
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-樣本制備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).
白細胞去除術(shù)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推薦介質(zhì)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免疫細胞療法參考資料：
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療法免疫細胞磁珠富集分選試劑盒產(chǎn)品資料由紅榮微再編輯整理。紅榮微再（上海）生物工程技術(shù)有限公司以“傳遞科學(xué)價值，服務(wù)科學(xué)研究”為宗旨，主營：干細胞、醫(yī)療、細胞治療、器官再生 四大板塊的產(chǎn)品。
選購19051 Stemcell EasySep™ Enrichment Kit CAR-T療法免疫細胞磁珠富集分選試劑盒歡迎您致電 華雅再生醫(yī)學(xué)旗艦公司：紅榮微再（上海）生物工程技術(shù)有限公司 ：152 1681 4001。華雅再生醫(yī)學(xué)-客服: 316 808 6348。
