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CellPore™ Transfection Kit 300

Transfection & intracellular delivery kit for cell engineering applications
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产品号 #100-1020_C

Transfection & intracellular delivery kit for cell engineering applications

产品优势

  • Efficiently deliver target cargo molecules into unactivated T cells, NK cells, PBMCs, and CD34+ HSPCs
  • Retain control over immune cell stimulation by avoiding electroporation-induced activation
  • Easily optimize delivery of cargo such as nucleic acids, gene editing complexes, proteins, and others in one experiment

产品组分包括

  • CellPore™ Transfection Kit 300 (Catalog #100-1020)
    • CellPore™ Delivery Medium, 2 x 2.5 mL
    • CellPore™ FITC-Dextran, 0.15 mL
    • CellPore™ Delivery Cartridges 300, 12 Cartridges/Bag
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CellPore™ Transfection Kit 300
Products for Your Protocol
To see all required products for your protocol, please consult the Protocols and Documentation.
  1. CellPore™ Transfection System
    CellPore™ Transfection System

    Cell mechanoporation system for transfection & intracellular delivery

概述
实验数据
产品说明书及文档
应用领域
相关材料与文献

概述

Efficiently deliver target cargoes to the cytosol of human unactivated immune and hematopoietic stem cells with the CellPore™ Transfection Kit 300. Designed for use with the CellPore™ Transfection System—a benchtop instrument for transfection and intracellular delivery through mechanoporation—the CellPore™ Transfection Kit 300 contains all the essential tools for conducting transfection reactions.

Our easy-to-use transfection system offers customization and flexibility for delivering a range of cargoes into hard-to-transfect cells with minimal cell perturbations. Use it to achieve superior viability and delivery compared to other cell transfection methods, all with minimal protocol adjustments and seamless integration into existing workflows.

The CellPore™ Transfection Kit 300 includes CellPore™ Delivery Cartridges for 20 - 200 μL reaction volumes, CellPore™ Delivery Medium for efficient cargo delivery and high cell viability, and CellPore™ FITC-Dextran as a positive control.

This kit is compatible for use with the following human cells:
• Unactivated Pan T cells
• Unactivated CD4 T cells
• Unactivated CD8 T cells
• Unactivated Regulatory T cells
• Unactivated Natural killer (NK) cells
• CD34+ hematopoietic stem and progenitor cells (HSPCs)
• Peripheral blood mononuclear cells (PBMCs)
This kit may also be compatible with other unactivated lymphocytes such as human B cells, human monocytes, and mouse T cells.

The kit supports two protocols. The CellPore™ Transfection Kit 300 delivery protocol makes it simple to optimize cargo delivery efficiency and maintain high cell viability by identifying the optimal pressure settings. Conduct optimization experiments tailored to your specific needs by following the detailed guidelines provided. Additionally, our CRISPR-Cas9 protocol is validated for editing unactivated primary human pan T cells, ensuring reliable performance for this application. Both protocols are available in the Product Information Sheets, located in the Protocols and Documentation section below.

For more information about CellPore™, including the intracellular delivery technology used in the CellPore™ Transfection System, visit the CellPore™ overview page.

Cell Type
T Cells
 
Species
Human
 
Application
Genome Editing
 
Area of Interest
Immunology
 

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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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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