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CellPore™转染试剂盒300

用于细胞工程应用的转染和细胞内递送试剂盒
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产品号 #（选择产品）

产品号 #100-1020_C

用于细胞工程应用的转染和细胞内递送试剂盒，可处理12个反应

产品优势

  • 高效地将目的物质分子递送到未激活的T细胞、NK细胞、PBMC和CD34+ HSPC中；
  • 通过避免电穿孔诱导的激活来控制对免疫细胞的刺激；
  • 在单次实验中轻松优化核酸，基因编辑复合物，蛋白质等目的物质的递送

产品组分包括

  • CellPore™转染试剂盒300（产品号 #100-1020）：
    • CellPore™递送液，2 × 2.5 mL
    • CellPore™ FITC-葡聚糖，0.15 mL
    • CellPore™递送管300，12个/袋
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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
相关产品

总览

使用CellPore™转染试剂盒300有效地将目的物质递送到未激活的人免疫和造血干细胞的细胞质中。CellPore™转染试剂盒300专为搭配通过机械穿孔进行转染和细胞内递送的CellPore™细胞转染系统使用而设计，包含进行转染反应的所有必需工具。

这套易于使用的转染系统提供定制空间和灵活性，将一系列物质递送到难以转染的细胞中的同时将对细胞的扰动降低到最低程度。使用CellPore™可以实现与其他细胞转染方法相比更好的活率和递送效率，不需要对现有的实验方法进行很大调整，可无缝整合到您现有的工作流程中。

CellPore™转染试剂盒300包括用于20 - 200 μL反应体积的CellPore™递送管，为高效货物递送和高细胞活率而设计的CellPore™递送液，以及作为阳性对照的CellPore™FITC-葡聚糖。

该试剂盒适用于以下人细胞：

•未活化的pan-T细胞

•未活化的CD4 T细胞

•未活化的CD8 T细胞

•未活化的调节性T细胞 •未活化的自然杀伤（NK）细胞 •CD34+造血干祖细胞（HSPC） •外周血单个核细胞（PBMC） 本试剂盒也可能与其他未激活的淋巴细胞类型，如人B细胞，人单核细胞和小鼠T细胞兼容。 该试剂盒可支持两套实验方案。CellPore™转染试剂盒300的递送优化实验方案可以简单地通过确定最佳的压力设定来优化目的物质递送效率并保持高细胞活率。根据您的具体需求，您可按照提供的详细指导进行优化实验。此外，我们的CRISPR-Cas9递送方案已被验证用于编辑未激活的原代人pan-T细胞，能够确保该应用的性能可靠。上述的两种实验方案都可以在产品说明书中找到，产品说明书位于页面下方的“Protocols and Documentations”部分。 关于CellPore™的更多信息，包括CellPore™细胞转染系统中使用的细胞内递送技术，请访问CellPore™总览页面。

细胞类型
T 细胞
 
种属
人
 
应用
基因组编辑
 
研究领域
免疫
 

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Data Figures

Optimizing Pressure Parameters for CellPore™ FITC-Dextran Delivery to Isolated Human Primary Cells

Figure 1. Optimizing Pressure Parameters for CellPore™ FITC-Dextran Delivery to Isolated Human Primary Cells

The graphs show the results of pressure sweeps to determine the best delivery parameters for CellPore™ FITC-Dextran in human (A) T and (B) NK cells from a leukopak and (C) cryopreserved human cord blood-derived CD34+ hematopoietic stem and progenitor cells (HSPCs). Delivery efficiency (orange bars) and cell viability (black circles) were assessed via flow cytometry on the day of delivery. Optimal delivery conditions were defined as those in which delivery efficiency is maximized while impact on cell viability is minimal. Control groups include untreated samples and endocytosis controls for comparison. The endocytosis control represents the natural uptake of CellPore™ FITC-Dextran in undelivered samples without pressure application. Data are presented as mean ± SD, with n = 3 - 18 replicates.

mCherry mRNA Delivery to Isolated Human Peripheral Blood NK Cells Using CellPore™

Figure 2. mCherry mRNA Delivery to Isolated Human Peripheral Blood NK Cells Using CellPore™

mCherry mRNA was delivered to freshly isolated human NK cells (2 x 106 cells in 20 µL reactions) from a leukopak using the CellPore™ Transfection System at 70 psi. Doses ranged from 0.2 to 4.0 µg of mRNA. mCherry expression was measured via flow cytometry 2 days after delivery. Control conditions included untreated NK cells and an endocytosis control (4.0 µg mRNA added without pressure application). Data shown as mean ± SD, n = 2.

The CellPore™ Transfection System Can Efficiently Knock Down Gene Expression in Unactivated Pan T Cells at a Wide Range of Cell Concentrations

Figure 3. The CellPore™ Transfection System Can Efficiently Knock Down Gene Expression in Unactivated Pan T Cells at a Wide Range of Cell Concentrations

Cas9 RNPs targeting B2M (A, B) or TRAC (C, D) genes were delivered to 0.5 - 25 x 106 isolated unactivated pan T cells in 50 µL reactions using the CellPore™ Transfection System at 90 psi. Flow cytometry was used to assess B2M (MHC-I) after 2 days or TRAC (TCRαβ) knockout efficiency after 6 days, alongside viability (using DRAQ7) in Pan T, CD4+, and CD8+ T cell subsets. Comparable viability and editing performance were observed across all tested cell numbers. Control groups included untreated T cells and scramble controls delivering non-targeting sgRNA Cas9 RNPs. Data presented as mean ± SD; n = 3.

The CellPore™ Transfection System Preserves Unactivated State of Pan T Cells

Figure 4. The CellPore™ Transfection System Preserves Unactivated State of Pan T Cells

Isolated unactivated pan T cells were manipulated using the CellPore™ Transfection System (delivery pressure 90 psi) or an optimized electroporation protocol. One and two days post-delivery, flow cytometry was used to assess CD69 expression, an early T cell activation marker. Electroporation increased CD69 expression in both (A) CD4+ and (B) CD8+ subsets of pan T cells. (C) Electroporation also elevated secretion of pro-inflammatory cytokines, measured using a bead-based multiplex cytokine assay 24 hours post-delivery. In contrast, the CellPore™ Transfection System maintained CD69 expression levels comparable to untreated control pan T cells. These findings underscore the gentle impact of the CellPore™ Transfection System on cellular function following intracellular cargo delivery. Data presented as mean ± SD; n = 3.

The CellPore™ Transfection System Enables Efficient Gene Knock-Out in CD34+ HSPCs

Figure 5. The CellPore™ Transfection System Enables Efficient Gene Knock-Out in CD34+ HSPCs

Cryopreserved cord blood-derived CD34+ HSPCs were cultured in complete StemSpan™ SFEM II medium for 24 hours. RNP complexes targeting the B2M or PTPRC gene were delivered to 5 x 104 HSPCs. After a culture period of 4 days, flow cytometry was used to determine viability and (A) B2M knock-out (assessed via MHC-I surface expression, representative histogram shown) or (B) PTPRC knock-out (assessed via CD45 surface expression, representative histogram shown). (C) A pressure sweep was performed to identify the optimal delivery pressure for achieving robust gene knock out in CD34+ HSPCs, which was determined to be 30 psi. (D) When utilizing the optimal 30 psi delivery pressure, the average editing efficiency for MHC-I knock-out was 91.8% ± 1.4, and 84.2% ± 2.7 for CD45 knock-out, while retaining high viability. Data are presented as mean ± SD, n = 2 - 7.

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

Document Type
Product Name
Catalog #
Lot #
Language
Document Type
Product Information Sheet
Product Name
CellPore™ Transfection Kit 300
Catalog #
100-1020
Lot #
All
Language
English
Document Type
Product Information Sheet - Special Applications Protocol
Product Name
CellPore™ Transfection Kit 300
Catalog #
100-1020
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
CellPore™ Transfection Kit 300
Catalog #
100-1020
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
CellPore™ Transfection Kit 300
Catalog #
100-1020
Lot #
All
Language
English

Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Research Area
Workflow Stages
Workflow Stages for Genome Editing
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Species Human
Legal Statement: The purchase of a CellPore™ Transfection System, CellPore™ Delivery Cartridges, and associated reagents (“CellPore™ Products”) conveys to the purchaser a limited, non-exclusive, non-transferable license to use CellPore™ Products, in accordance with STEMCELL’s Terms and Conditions of Sale (www.stemcell.com/terms-and-conditions-general) and all applicable laws and regulations, and solely for research-use-only applications (which applications exclude any commercial application or any therapeutic, prophylactic, diagnostic application, as well as any development and/or commercialization of products therewith or developed therefrom). STEMCELL or its licensor(s) grant no additional license rights other than those explicitly granted herein, and except for such limited license rights, all other intellectual property and proprietary rights in and to CellPore™ Products are and shall remain the exclusive property of STEMCELL and its licensors. The purchaser of CellPore™ Products agrees to prevent unauthorized use, access, copying, or disclosure of any intellectual property comprised in CellPore™ Products. The purchaser shall not, and shall not permit anyone to, copy, create any derivative work, reverse engineer, disassemble, decompile, or make improvements to CellPore™ products. Users and purchasers of CellPore™ products hereby grant to STEMCELL exclusive ownership and all rights, title, and interest in and to any improvements made, by purchasers or users, to the CellPore™ products (including any improvements by purchasers or users to the intellectual property directly related to CellPore™ products), provided however that the user or purchaser who makes the improvement retains a limited, non-transferable license to use such improvement for its internal, non-commercial research use, in accordance with this agreement, only so long as such user or purchaser is in compliance with this agreement. All data, information, and results (to the extent they do not constitute improvements of CellPore™ products) entered, stored, compiled, generated, and/or analyzed by purchasers of CellPore™ products using the CellPore™ products, in accordance with all of the foregoing, shall be the property of such purchaser. CellPore™ Transfection System and CellPore™ Delivery Cartridges (“CellPore™ Products”) are protected by patents in the U.S. and elsewhere. More information can be found at www.cellpore.com. The list of products and patents contained therein are subject to change as a result of routine prosecution and dependent on the patent life cycle as determined by the laws of each jurisdiction in which a relevant patent is in effect. UNLESS OTHERWISE STATED, PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES. FOR PRODUCT-SPECIFIC COMPLIANCE AND INTENDED USE INFORMATION, REFER TO THE PRODUCT INFORMATION SHEET. GENERAL INFORMATION ON QUALITY AT STEMCELL MAY BE FOUND AT WWW.STEMCELL.COM/COMPLIANCE.
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