Can frozen PBMCs be used as the starting population for NK cell expansion with Catalog #s CLD004 and CCM032?

Frozen PBMCs work well as a starting population for subsequent NK cell expansion using CLD004. Cells should be thawed and washed twice at 300 x g for 5 minutes, to remove cryoprotectant. Gentle mixing is recommended. Allow the cells to rest for 1-4 hours in a 37 C incubator prior to expansion.

Can serum-free media be used to expand NK cells in the Cloudz Human NK Cell Expansion Kit, Catalog # CLD004?

Yes. We have found a robust expansion of NK cells using Bio-Techne brand ExCellerate Human NK Cell Expansion Media, Xeno-Free, Catalog # CCM032. We have found that this medium is a superior alternative to other serum-free medias on the market.

Have you tried expanding purified NK cells instead of PBMCs using CCM032?

Yes, we have tried expanding purified NK cells using the same protocol, and it works well.

Is the formulation of the media used in CellXVivo Human NK Cell Expansion Kit, catalog # CDK015, different than Human NK Cell Expansion Media, catalog # CCM032?

They do not have the same formulation. The expansion medium used in CDK015 is not serum-free. It contains human serum.

What is the expected viability of cells after 9 days of culture, when following the Cloudz Human NK Cell Expansion Kit protocol with ExCellerate™ Human NK Cell Expansion Media, Xeno-Free media, catalog # CCM032?

The expected cell viability is 70-90%.

ExCellerate™ Human NK Cell Expansion Media, Xeno-Free

Name: ExCellerate™ Human NK Cell Expansion Media, Xeno-Free
Brand: R&D Systems
SKU: CCM032
Rating: 3 (1 reviews)

R&D Systems, part of Bio-Techne | Catalog # CCM032

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Product Datasheet / COA / SDS

New Version Available - CCM037

For robust NK cell expansion under xeno-free conditions.

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Product Summary
Scientific Data
Kit Contents
Preparation & Storage
Product Documents
Specific Notices

Key Product Details

Features

Media for the ex vivo expansion and culture of human Natural Killer lymphocytes under xeno-free conditions.

Key Benefits

Robust NK cell expansion under serum-free and xeno-free conditions
Compatible with a variety of NK cell activation methods
Supports NK cell expansion from PBMCs or purified CD3⁺-depleted PBMCs

Species

Human

Product Summary for ExCellerate™ Human NK Cell Expansion Media, Xeno-Free

Why Use ExCellerate™ Human NK Cell Expansion Media?

Natural Killer (NK) cells play an important role in both the adaptive and innate immune responses that control infection, autoimmunity, and tumor immunosurveillance. Due to their intrinsic and non-specific anti-tumor activity, human NK cells have been employed as anti-cancer therapies. Robust expansion and maintenance of NK Cells is critical to facilitate NK cell therapy.

Versatility of ExCellerate Human NK Cell Expansion Media

ExCellerate Human NK Cell Expansion Media is optimized for NK cell expansion when used in combination with Recombinant Human Cytokines from R&D Systems. However, this versatile media also provides robust growth using a variety of cytokine combinations (IL-2, IL-12, IL-18, and IL-21) and cell activation methods, including particle, bead-based, or plate-bound Human NKp46/NCR1 Antibody. The activation and cytokine/growth factor combinations used with this media should be optimized by application or experimental protocol.

Scientific Data Images for ExCellerate™ Human NK Cell Expansion Media, Xeno-Free

NK Expansion Protocol.

Fourteen day protocol for the expansion of human NK cells using ExCellerate Human NK Cell Expansion Media (Catalog # CCM032).

Flow Cytometry Analysis of Expanded Human NK Cells.

Human PBMCs were expanded in vitro for 14 days. NK Cells were collected on days 0, 8, 11, and 14 of expansion, and stained with Human CD3 Fluorescent-conjugated antibody (FAB100F) and Human NCAM-1/CD56APC-conjugate antibody, and analyzed by flow cytometry. Compared to unexpanded PBMCs on Day 0, Days 8, 11, and 14 show an increased number of CD3- CD56+ NK Cells. Flow quadrants were set based on isotope-stained samples.

NK Cell Expansion and Purity Following Expansion.

Human PBMCs were expanded in vitro for 14 days. Cells were evaluated at Days 0, 8, 11, and 14 for fold expansion (A) and population characterization (B). Following 14 days of expansion, NK Cells showed approximately 500-fold expansion (A) and a purity of 70.7% of CD3- CD56+ NK Cells (B).

Kit Contents for ExCellerate™ Human NK Cell Expansion Media, Xeno-Free

500 mL of ExCellerate Human NK Cell Expansion Media

Formulation, Preparation, and Storage

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the recommended temperature.

Storage

Store the unopened product at -20 °C. Do not use past expiration date.

Product Documents for ExCellerate™ Human NK Cell Expansion Media, Xeno-Free

Product Datasheets

Download Product Datasheets

COA

SDS

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Product Specific Notices for ExCellerate™ Human NK Cell Expansion Media, Xeno-Free

FOR LABORATORY RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.

The safety and efficacy of this product in diagnostic or other clinical uses has not been established.
This reagent should not be used beyond the expiration date indicated on the label.
Results may vary due to variations among cells derived from different donors.

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