Cloudz™ Cell Activation Kits

Cloudz for Cell Activation and Expansion
The Sky is the Limit with Cloudz

Robust and reproducible cell activation and/or expansion is necessary for successful cell culture and is a critical component of cell therapy manufacturing processes. R&D Systems is proud to provide a unique and customizable platform to support both discovery stage and clinical scale programs.

 

What Are Cloudz Reagents?

Cloudz kits are used in the expansion and activation of immune cells for research & manufacturing.

Cloudz reagents are microspheres composed of an alginate-based hydrogel. The microspheres can be functionalized with a variety of ligands including antibodies, proteins, and small molecules to activate desired cells and enable cell expansion. Exposure of immune cells to the ligands on Cloudz microspheres activates intracellular signaling to promote cell expansion and provide you with a robust cell culture.

The alginate hydrogel microspheres quickly dissolve when exposed to our proprietary Release Buffer supplied in the kits, allowing for gentle removal of the microspheres. This dissolving property also allows you to precisely and rapidly remove residual Cloudz and their corresponding ligand from the expanded cells of interest.

 

What Are the Benefits of Cloudz Cell Activation and Expansion Reagents?

  • Efficient: Release buffer dissolves microspheres rapidly without a magnetic-based removal step
  • Customizable: Choose a ligand of your choice to activate your desired cell type
  • Gentle: The microspheres gently activate your cells to maximize expansion
  • Flexible: Cloudz can work within many expansion platforms to suit your existing process
  • Optimized for use with R&D Systems ExCellerate™ media & GMP cytokines and Wilson Wolf’s G-Rex cell culture vessels.

Cloudz kits are used for cell expansion and cell activation of immune cells.

 

Cloudz for Immune Cell Therapy Manufacturing

A reliable and scalable cell expansion platform is critical for a successful growing process. Research-and GMP-grade Cloudz kits, as well as custom options, are available to support consistent and seamless process translations. For our complete immune cell therapy manufacturing solution, visit ScaleReady.

 

Cloudz Expansion Protocol – A Simple 3-step Process

  1. Add Cloudz reagent to your cell culture.
  2. Incubate cell culture with Cloudz for at least 2 days.
  3. Add release buffer to dissolve Cloudz microspheres.

Expanded cells are ready for use, free of residual particulates.

NK Cells are expanded using the Cloudz Human NK Cell Expansion Kit. After 10 days in culture, Human PBMCs achieved an expansion rate of 300-fold expansion and a purity of 94.4% CD3-CD56+ cells.

Fig. 1. NK Cell Expansion and Purity Following Expansion using the Cloudz Human NK Cell Expansion Kit. Human PBMCs were expanded in vitro for 10 days using the protocol and reagents included in the Cloudz Human NK Cell Expansion Kit. Cells were evaluated at Days 6, 7, 9, and 10 for fold expansion (A) and population characterization (B). Following 10 days of expansion NK cells showed approximately 300-fold expansion (A) and a purity of 94.4% of CD3-CD56+ NK cells (B).

 

Cloudz Cell Activation Kits

Human T Cell Activation Kits

The Cloudz Human T Cell Activation Kits support the expansion of human T cells and the stimulation of human peripheral blood lymphocytes. The CD3 and CD28 antibodies on these Cloudz reagents were selected to optimally activate and expand CD4+ and CD8+ T cells. Activation of T cells with Cloudz microspheres enables efficient cell expansion while maintaining a high population of naïve-like memory cells. This kit is available in both research use only and GMP grades.  

Description Expansion Capacity Format Ligands Optimized for use with
GMP Cloudz Human T Cell Activation Kits > 50-fold expansion of T cells from PBMCs in 10 days GMP Fully Humanized CD3 and CD28 Antibodies

ExCellerate Human T Cell Expansion Media, Xeno-Free

GMP ProDots Proteins

R&D Systems GMP Cytokines

Cloudz Human T Cell Activation Kits > 50-fold expansion of T cells from PBMCs or purified T cells in 10 days Research Fully Humanized CD3 and CD28 Antibodies

ExCellerate Human T Cell Expansion Media, Xeno-Free

Recombinant Human IL-2

Recombinant Human IL-7

Recombinant Human IL-15

 

Human Regulatory T Cells (Treg) Expansion Kits

Regulatory T cells, also known as suppressor cells, are a subset of CD4+ T cells that maintain immune homeostasis by suppressing immune responses and controlling autoimmune recognition. The Cloudz Human Treg Expansion Kits provides robust expansion of FoxP3+ human Treg cells from a population of CD4+ T cells.

Description Expansion Capacity Format Ligands Optimized for use with
GMP Cloudz Human Treg Cell Activation Kit 500-fold expansion of Treg cells from CD4+ T cells in 9 days GMP Fully Humanized CD3 and CD28 Antibodies

ExCellerate Human T Cell Expansion Media, Xeno-Free

GMP ProDots Protein Recombinant Human IL-2

Cloudz Human Treg Cell Activation Kit 500-fold expansion of Treg cells from CD4+ T cells in 9 days Research Fully Humanized CD3 and CD28 Antibodies

ExCellerate Human T Cell Expansion Media, Xeno-Free

Recombinant Human IL-2

 

Human Natural Killer (NK) Cell Expansion Kit

Natural killer cells are innate lymphoid cells with intrinsic cytotoxic and cytokine producing capabilities. These Cloudz kits provide a feeder-free expansion protocol that selectively expands NK cells from peripheral blood mononuclear cells (PBMCs) or purified NK cells. The CD2 and NKp46 antibodies provide key activating signals for NK cells. The Cloudz kit containing humanized antibodies is designed to streamline translation to clinical manufacturing processes.  

Description Expansion Capacity Format Ligands Optimized for use with
Cloudz Human NK Cell Expansion Kit Up to 200-fold expansion of NK cells in 10 days Research Mouse CD2 and NKp46 Antibodies

ExCellerate Human NK Cell Expansion Media, Xeno-Free

Recombinant Human IL-2

Recombinant Human IL-12

Recombinant Human IL-18

Recombinant Human IL-21

Cloudz Human NK Cell Expansion Kit (Coming Soon) Up to 200-fold expansion of NK cells in 10 days Research Fully Humanized CD2 and NKp46 Antibodies

ExCellerate Human NK Cell Expansion Media, Xeno-Free

Recombinant Human IL-2

Recombinant Human IL-12

Recombinant Human IL-18

Recombinant Human IL-21

 

Design and Build Your Own Screening Cloudz (Coming Soon!)

Screening Cloudz are derivatized with streptavidin which offers you the flexibility to conjugate them with any biotinylated ligand such as our precisely labeled Avi-tag biotinylated proteins. With Screening Cloudz you can easily evaluate multiple ligands for the best cell activation or expansion in your particular assay. Once you have identified the optimum combination of ligands, we can manufacture a Cloudz kit at the scale you need. 

Ligands are typically chosen based on their ability to provide activating signals to the target cell. This includes antibodies to cell surface molecules and ligands that bind cell surface receptors. Inhibitory signals can be delivered by targeting inhibitory receptors or by using Cloudz derivatized with blocking antibodies or antagonistic ligands.

We offer an extensive portfolio of premium reagents including recombinant proteins, antibodies, peptides, and small molecules. Our Custom Protein Conjugation & Biotinylation services and Custom Chemistry Services can modify the molecules of your choice.

Book an appointment with a Cell and Gene Therapy Specialist to discuss the possibility of using Screening Cloudz in your application.

 

Not Seeing the Ligands You Need? Let Us Customize Your Cloudz.

Our Custom Services team will work with you to produce it. We have access to all Bio-Techne catalog products as well as hundreds of non-catalog reagents, and we can also custom design ligands for your optimum Cloudz kit. When you are ready to take the step to GMP-grade materials, we can deliver that too. Contact our Custom Team to learn more.

 

ScaleReady

Bio-Techne believes in the power of partnerships. We have joined with Fresenius Kabi and Wilson Wolf to bring you ScaleReady™ for immune cell therapy manufacturing.

ScaleReady brings cell and gene therapies to life by streamlining manufacturing processes across cell culture, cell activation and expansion, gene editing, and cell processing.

Learn More at ScaleReady.com

 

 

Frequently Asked Questions

Find out the answers to some of your frequently asked questions about Cloudz.

Are the Cloudz Cell Activation/Expansion Kits cell isolation kits?

No, our Cloudz kits do not isolate cells but are cell activation/expansion products. However, we offer T Cell Enrichment Columns or MagCellect™ Cell Selection Kits for cell isolation.

What is the difference between cell activation & cell expansion?

Exposure of immune cells to the ligands on Cloudz microspheres activates intracellular signaling to promote cell expansion and provide you with a robust cell culture during activation and expansion steps of cell culture.

Why are flasks placed vertically instead of horizontally in the protocol for Cloudz NK Cell Expansion kit?

The vertical positioning is mostly important for the first 3-5 days of culture (depending on donor). It maintains a higher cell density and allows them to survive to proliferate at later timepoints.

Is the expansion rate of cells independent of the source of the cells?

Donor to donor variability plays a significant role in the growth kinetics of cells, and will impact the ultimate expansion and phenotype of your expanded cells.

When should I remove my Cloudz particles from my cells?

The Cloudz microspheres can either be dissolved after 48 hrs, or remain for the entire culture period

How do I remove Cloudz microspheres from my cell culture when my cells have reached maximal confluency?

The Release Buffer in the Kit can be added to culture to completely dissolve away Cloudz Microspheres. Cells can then be spun down and washed to completely clear the release buffer from culture. Do not leave cells in release buffer for more than 30 minutes.

How do I confirm removal of Cloudz microspheres from my culture?

Cloudz Microspheres can be assessed using the Micro-Flow Image-based particle analysis instrument to observe dissolution of the Cloudz.

Should I use serum-free media or serum-containing media for expansion of NK cells in the Cloudz Human NK Cell Expansion Kit?

Expansion of NK Cells in serum-free media is more challenging than expansion in media containing serum. Comparatively, donor to donor variability has a larger impact on NK cell expansion in serum-free media than in serum-containing media. We have found a robust expansion of NK cells using Bio-Techne brand ExCellerate Human NK Cell Expansion, Xeno-Free Media.

Cloudz Human T Cell Activation Kit and the Cloudz Human Treg Expansion kit both contain Cloudz CD3/CD28 antibodies. Can the antibodies provided in the kits be substituted for various cell types and provide the same results?

While both kits have the similar antibodies, each product is manufactured and optimized to each type of cell expansion (T cell or Treg). Use of Cloudz CD3/CD28 from one kit to another is not supported.