EasySep™人T細胞分選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合物(Tetrameric Antibody Complexes)和dextran包被的磁珠標記后移除。被標記細胞不需要過柱,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CAR-T治療下游細胞培養(yǎng)。
EasySep™人T細胞分選試劑盒是EasySep™人T細胞富集試劑盒的新版(貨號:19051),細胞分選更快速,短至8分鐘即可完成免細胞磁珠分選。
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進行磁珠分選篩選T細胞。Stemcell的EasySep™人T細胞分選試劑盒可以用于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細胞分選試劑盒的磁珠分選采用負選方式,可以避免標記T細胞,以防標記抗體對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細胞分選試劑盒
英文名稱 EasySep™ Human T Cell Isolation Kit
用于標記1 x 109個細胞。
EasySep™人T細胞分選試劑盒是套裝,包含:
EasySep™ Human T Cell Isolation Cocktail(1mL) T細胞分選抗體復合物
EasySep™ Dextran RapidSpheres™ (1mL) 磁珠
貨號:17951
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17951 Stemcell EasySep Human T Cell Isolation Kit CAR-T治療人T細胞磁珠分選試劑盒歡迎您致電 華雅再生醫(yī)學旗艦公司: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-客服: 316 808 6348。
EasySep人T細胞分選試劑盒:適用樣本
EasySep™人T細胞分選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。目標細胞被抗體復合物和磁珠標記,以進行負選。
諾華的CAR-T免疫細胞療法使用同樣的樣本1。
EasySep人T細胞分選試劑盒應(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(免疫細胞磁珠分選法,EasySep™人T細胞分選試劑盒)——CAR-T細胞活化、擴增和基因編輯——CAR-T細胞回輸
其他應(yīng)用
流式細胞術(shù)、細胞培養(yǎng)(CAR-T免疫療法)或DNA/RNA提取
EasySep Human T Cell Isolation Kit組份
成分 | 貨號 | 數(shù)量 | 保存 | 保質(zhì)期 | 內(nèi)容 |
EasySep™ Human T Cell Isolation Cocktail EasySep™分選抗體混合物 | 17951C | 1 x 1 mL | 保存 2 - 8°C,不能冷凍 | 效期內(nèi)穩(wěn)定 | 單克隆抗體復合物(加PBS) |
EasySep™ Dextran RapidSpheres™ | 50103 | 1 x 1 mL | 保存 2 - 8°C,不能冷凍 | 效期內(nèi)穩(wěn)定 | 磁珠懸液(水) |
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.
EasySep人T細胞分選試劑盒:磁珠分選常規(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™ RapidSpheres™ 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。
EasySep人T細胞磁珠分選試劑盒優(yōu)點
方便快捷,新版短至8分鐘。無柱分選
純度zui高達98%
細胞保持Untouched
T細胞負選方案介紹
在負選時,非目標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)標記的細胞
若將抗體結(jié)合到目標細胞上會造成抑制或活化,影響下游研究
目標細胞沒有特異性抗體可用來標記
需要去除某些類型的細胞
文獻——使用Stemcell EasySep人T細胞分選試劑盒進行CAR-T療法的磁珠分選
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 | 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 | 17951 17951RF |
EasySep Human T Cell Isolation Kit免疫細胞磁珠分選:相關(guān)產(chǎn)品
品牌 | 貨號 | 產(chǎn)品描述 |
Stemcell | 19051 | EasySep™ Human T Cell Enrichment Kit,EasySep™人T細胞分選試劑盒的舊版 |
Stemcell | 18000 | EasySep™磁極 |
Stemcell | 20144 | EasySep™ Buffer |
概要EasySep Human T Cell Isolation Kit
The EasySep™ Human T Cell Isolation 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細胞分選試劑盒說明書
描述EasySep Human T Cell Isolation Kit Description
Isolate untouched and highly purified T cells from fresh or previously frozen human peripheral blood mononuclear cells (PBMCs) or washed leukapheresis samples in as little as 8 minutes by immunomagnetic negative selection.
– Fast, easy-to-use and column-free
– Up to 98% purity with high recovery
– Untouched, viable cells
This 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 Isolation Kit-樣本制備Sample Preparation
For available fresh and frozen samples, see www.stemcell。。com/primarycells.
PERIPHERAL BLOOD
Prepare a PBMC suspension from whole 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)
Wash the peripheral blood leukapheresis sample by adding an equivalent volume of recommended medium or PBS containing 2% fetal bovine serum (FBS).
Centrifuge at 500 x g for 10 minutes at room temperature (15 - 25°C). Remove the supernatant and resuspend the cells at 5 x 10^7 cells/mL in recommended medium.
EasySep Human T Cell Isolation 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.
17951 Stemcell EasySep Human T Cell Isolation Kit CAR-T治療人T細胞磁珠分選試劑盒產(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“傳遞科學價值,服務(wù)科學研究”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17951 Stemcell EasySep Human T Cell Isolation Kit CAR-T治療人T細胞磁珠分選試劑盒歡迎您致電 華雅再生醫(yī)學旗艦公司: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-客服: 316 808 6348。