Streamline Your Large-Scale Cell-Based Research Workflows

This blog post was written by STEMCELL Technologies, who provide cell isolation products, specialized cell culture media, primary cells, and supporting reagents for use in life sciences research across the basic to translational research continuum. STEMCELL Technologies’ in-house Contract Assay Services (CAS) work with you to design and perform in vitro potency and safety studies for drug discovery and development. Their services are available on the Scientist.com marketplace.

Developing workflows for large-scale biomedical research, including drug discovery and the development of cell-based vaccines and therapeutics, requires significantly larger starting biological materials than conventional research methodologies. Thus, large-scale research strategies require a focus on advancing the collection, processing and analysis of large quantities of specific cell types for downstream applications. As researchers strive toward standardized and scalable workflows, the results can help them understand complex cellular phenomena more effectively and allow them a quicker transition from bench to clinic.

To meet these growing demands of scalable workflows, scientists must consider strategies for increasing the number of cells being sourced, isolated and cultured to streamline their research. Choosing large-scale human primary cell sources such as peripheral blood-sourced leukopaks or whole blood as starting sources is one key step in that direction. Another key step to increase efficiency is to scale up cell separation workflows so that a large number of cells can be isolated from one or multiple sources in the fewest steps. Large-scale cell isolation can accelerate drug discovery and immunotherapy development by enabling researchers to spend less time isolating cells and more time on their downstream experiments. Efficient cell-based research depends not just on working with a reliable primary cell supplier, but also on ensuring workflows that are standardized and scalable. STEMCELL Technologies can support your cell-based research workflows by providing high-quality, ethically sourced human primary cells, along with the most efficient isolation technologies and quality cell culture and expansion media, supplements, reagents and protocols.

Additional benefits of large-scale research workflows include:

• Reduced biological variability of results: Using cells isolated from the same donor instead of multiple donors reduces biological variability. In the context of drug discovery and finding therapy candidates, using the same donor ensures that the obtained results are due to the therapy candidate and not due to biological differences in donors.
• Fewer donors required: Isolating large numbers of cells from specific donors reduces the need to keep searching for more donors who meet the desired criteria.
• Fewer isolations needed when establishing cell banks: Obtaining sufficient cells to bank for future experiments avoids the need to repeat cell isolations later on.

Scalable Cell Sourcing

Human primary cells have already become an efficient and integral tool for researchers aiming to reduce extensive in vivo validation and facilitate their translation of basic research into preclinical or clinical applications. Scale up your primary cell-based research by using large-scale peripheral blood sources such as whole blood or leukopaks. These single-donor cell sources increase the physiological relevance of cell culture systems, enabling you to generate meaningful data that is more predictive of in vivo outcomes.

Get Quality Fresh Cells for Your Research from Whole Peripheral Blood*

Peripheral whole blood samples contain all blood cell types: i.e., erythrocytes (red blood cells), leukocytes (white blood cells) and thrombocytes (platelets), suspended in blood plasma. One of the advantages of using whole blood as a cell source is the freshness of the material, which is essential for studying sensitive cells like neutrophils. However, as isolating peripheral blood mononuclear cells (PBMCs) from whole blood can be tedious, a buffy coat is a widely used first-step separation product with a higher PBMC concentration than whole blood. The generation of a buffy coat takes advantage of the specific density differences among the various cell types within blood. STEMCELL provides whole peripheral blood in both small volumes (< 100 mL) and large volumes (≥ 450 mL), collected and supplied in 10 mL Vacutainer® tubes and collection bags, respectively, using acid citrate dextrose solution A (ACDA), ethylenediaminetetraacetic acid (EDTA) or sodium heparin (Na heparin) as an anticoagulant. Refer to Table 1 below for a comparison of large-volume samples.

Get the Cells You Need from Fresh or Frozen Peripheral Blood Leukopaks

Leukopaks are enriched leukapheresis** products containing a variety of immune cells, including T cells, B cells, natural killer (NK) cells, etc. Compared to whole blood and buffy coat, a full-size leukopak contains much higher numbers of leukocytes per volume — typically greater than 9 billion cells in an average volume of < 200 mL. This makes leukopaks an ideal source of human primary cells when large numbers of cells are required from a single donor. Compared to other cell sources, such as whole blood and PBMCs, leukopaks also offer higher yields of multiple immune cell types, standardization of assays and flexibility in experiments due to long-term storage capabilities.

Mobilized Leukopaks*
Mobilized leukopaks are peripheral blood leukopaks mobilized with G-CSF (Filgrastim or NEUPOGEN®), plerixafor (Mozobil®) or a combination of both. These cell sources contain large numbers of single-donor CD34+ hematopoietic stem and progenitor cells (HSPCs), thereby helping you minimize donor-to-donor single-cell variability and ensuring consistency across your HSPC studies. At STEMCELL, leukopaks mobilized with G-CSF, Plerixafor or a combination of G-CSF and Plerixafor (Combo) are available in full sizes and different collections based on the mobilization regimen.

Fresh Leukopaks (Non-Mobilized)*
Fresh leukopaks are typically used to study the functions of white blood cells within a short period of time after collection. This is because fresh leukopaks contain viable and functional white blood cells that can be used immediately after collection without any loss of viability or functionality. Fresh leukopaks are optimal for experiments requiring high cell functionality or sensitive cell types such as NK cells and dendritic cells. Fresh leukopaks are available from STEMCELL in full, half, quarter and tenth sizes.

Frozen or Cryopreserved Leukopaks (Non-Mobilized)
Frozen leukopaks, as the name suggests, are leukopaks that are cryopreserved and stored at a subzero temperature (normally ≤ -135°C) to preserve the biological sample for long-term storage. Frozen leukopaks are ideal for assays requiring a large number of cells or experiments conducted over extended periods of time. At STEMCELL, frozen leukopaks are processed shortly after collection, following stringent protocols, and cryopreserved in CryoStor® CS10 — preserving quality and ensuring optimal cell viability and purity post-thaw. Frozen leukopaks are available in full, half, quarter and tenth sizes. While using frozen leukopaks, it is critical to preserve cell functionality during thawing, by either optimizing your cell thawing protocol or using ThawSTAR® CB Automated Thawing System — reducing the risk of human error and standardizing cell thawing for scalable and reproducible results. Refer to Figure 1 below for a representative chart showing the average frequencies of major immune subsets in a frozen leukopak.

Looking for insights and techniques for efficient leukopak sample processing? Watch the leukopak processing webinar and reduce the time and reagents required to process cells of interest.

Scalable Immunomagnetic Cell Isolation

Isolating cells from large-volume samples such as leukopaks or whole blood is a common procedure that precedes many immunology studies. Working with large volumes can be challenging, as the sample often needs to be split and processed in parallel, which can be laborious and may delay downstream studies. Scaling up, in the context of cell isolation, involves transitioning to a platform that supports the processing and isolation of desired cells from large-volume samples to reduce the number of steps involved.

Platforms that harness immunomagnetic cell isolation technology, such as EasySep™ — a fast, easy-to-use and column-free magnetic system — can provide an efficient, scalable method of isolating highly purified cells from both small and large-volume samples that are ready for downstream applications.

When using such platforms, the first step involves processing the large-volume sample into a single-cell suspension. The method you choose to prepare the single-cell suspension will depend on the cell type you want to isolate. Once you have the single-cell suspension, using a large-scale approach enables you to isolate a large number of desired cells in a single cell separation procedure. In contrast, the small-scale approach typically requires multiple rounds of cell isolation.

Speed Up Cell Isolation with the Easy 250 EasySep™ Magnet
The Easy 250 EasySep™ Magnet simplifies cell separation when processing sample volumes of up to 225 mL, and is based on column-free immunomagnetic EasySep™ cell isolation technology. Instead of splitting your cell suspension and performing multiple rounds of cell isolation, you can process samples of up to 225 mL and 1.25 x 10¹⁰ cells in as little as 20 minutes with a single isolation. The Easy 250 EasySep™ Magnet generates a magnetic field in the interior cavity that is strong enough to separate cells labeled with magnetic EasySep™ particles. With this design, the labeled cells are pulled to the sides of the tube when the sample is placed in the magnet, while untouched cells can be pipetted off into a new tube. The Easy 250 EasySep™ Magnet is designed for use with a standard T-75 cm² cell culture flask and EasySep™ reagents to speed up your cell isolations from large-volume samples such as full-size leukopaks and whole blood.

Learn the steps and key considerations on how the Easy 250 EasySep™ Magnet simplifies cell separation when processing large-volume samples by watching this virtual poster presentation.

How Does the Easy 250 EasySep™ Magnet Perform?

The Easy 250 EasySep™ Magnet and compatible EasySep™ kits are designed for the isolation of the following cell types as part of your cell processing workflow. Obtain purities of up to 99% for human cell types isolated from large sample volumes (as shown in Figure 2 below), including:

• T cells and T cell subsets
• B cells
• Monocytes
• Natural killer cells
• Peripheral blood mononuclear cells
• And more!

You can explore more data to see how the Easy 250 EasySep™ Magnet performs when isolating different cell types. STEMCELL also provides a wide range of large-scale EasySep™ kits compatible with the Easy 250 EasySep™ Magnet for isolating your cells of interest using both negative and positive cell isolation protocols.

Scalable Workflow Solutions

To ensure optimal activation, expansion and differentiation of cells for specific downstream applications, STEMCELL also offers ImmunoCult™ cell culture reagents as well as cytokines and recombinant proteins. The ImmunoCult™ cell culture media, activators and supplements allow you to culture various cell types, including T cells, B cells, NK cells, dendritic cells and macrophages, under defined stimulatory conditions for consistent and reliable results. You can streamline your T cell therapy development with our new high-performance ImmunoCult-XF media and GMP ImmunoCult T Cell Activators, optimized for use in T cell therapy development and manufacturing, providing excellent T cell growth and viability without the use of magnetic beads, feeder cells or antigens. Achieve high-quality cultures and reproducible results with premium cytokines, chemokines and growth factors. Choose from more than 200 reagents supporting a wide variety of applications, including immunology and stem cell research, with multiple sizes and animal component-free options available.

STEMCELL Technologies offers a variety of high-quality, ethically sourced human primary cells* and efficient EasySep™ cell separation technology to meet your research needs. We can also accommodate your unique requirements with personalized service and customizable products, including products that meet specific isolation and anticoagulant requirements, custom cell isolation kits, cell culture media and more. To learn more, visit www.stemcell.com.

*Certain products are only available in select territories.
** Leukapheresis is a process of collecting white blood cells (WBCs) from peripheral blood of a healthy donor and returning the remainder to the circulation.

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