Automate Your PBMC Isolations
with RoboSep™
Considering an Alternative Way to Isolate PBMCs?
Peripheral blood mononuclear cells (PBMCs) are of increasing popularity in clinical research as they have several applications, including cell and gene therapy development, biomarker identification, and research into immune responses to treatment. While density gradient centrifugation and magnetic cell isolation are two methods that are commonly used to isolate PBMCs from whole blood, performing these techniques manually can be laborious, time-consuming, and leave room for error, which can delay downstream applications and research progress.
One way to overcome these hurdles is by automating PBMC isolation with platforms that enable you to reduce hands-on time and increase sample throughput. Automating PBMC isolation can streamline your workflows, save hands-on time, and progress your research faster.
Benefits of automating PBMC isolation include:
- Faster and More Efficient isolation than manual methods. This is beneficial for large-scale studies or clinical trials, where time and resources are often limited.
- More Consistent isolation, as they rely on programmed protocols. In contrast, manual techniques can be subject to human error.
- Reduced Variability that comes with manual isolation processes, such as the dilution, stratification, and collection of PBMCs from the density gradient.
- Reduced Risk of Contamination, which can occur during manual isolation and purification. This is important for maintaining the integrity of research samples and ensuring accurate and reliable results.
Ultimately, automation provides solutions for obtaining a large number of highly purified PBMCs in an efficient manner, to promote high-quality results and advancements in your research. Our RoboSep™ instruments seamlessly integrate the reliability, convenience, and throughput of a liquid-handler with the flexibility, speed, and fidelity of an immunomagnetic cell isolation system. RoboSep™-S and RoboSep™-16 perform all sample labeling and magnetic separation steps, thereby increasing laboratory output and efficiency while minimizing sample handling and technician time.
Why Automate PBMC Isolation Using RoboSep™?
- Minimize hands-on time and increase sample throughput
- Obtain highly purified mononuclear cells with improved RBC, granulocyte, and platelet clearance compared to density gradient centrifugation
- Load your samples and reagents, press “Run,” and come back to your isolated cells
- Isolate cells from cord blood, bone marrow, buffy coats, leukapheresis products, and leukocyte reduction system (LRS) cones
Automating PBMC Isolations Using RoboSep-S™
RoboSep™-S offers true walk-away automation of immunomagnetic PBMC separation. Using the EasySep™ Direct Human PBMC Isolation Kit, PBMCs are separated based on cell surface markers, rather than cell density, within the RoboSep™-S instrument. This results in highly purified PBMCs that are ready for downstream applications, all while reducing hands-on time, eliminating cross-contamination, and increasing sample processing throughput.
How Does the RoboSep™-S Instrument Work?
Table 1. Automated PBMC Isolation Using RoboSep™-S Reduces the Overall Cell Separation Time Compared to Density Gradient Centrifugation
| Step | Duration (minutes) | ||
|---|---|---|---|
| Manual Density Gradient Centrifugation |
Manual EasySep™ Direct |
Automated EasySep™ Direct on RoboSep™-S |
|
| Preparation of tubes with density gradient medium and layering sample | 7 | - | - |
| Loading samples and reagents onto the RoboSep™-S instrument | - | - | 5 |
| Cell isolation | 45 | 20 | 30 |
| Cell collection | 10 | 1 | 1 |
| Optional: Wash step to remove platelets | 15 | - | - |
| Optional: Red blood cell lysis | 15 | - | - |
| Final centrifugation step | 15 | Optional | Optional |
| Total time for cell separation | ≥ 77 Minutes | ~ 21 Minutes | ~ 36 Minutes |
| Total hands-on time | ≥ 32 Minutes | ~ 21 Minutes | ~ 6 Minutes |
Steps and approximate times to isolate monononuclear cells from whole blood samples when following using density gradient centrifugation or EasySep™ Direct Human PBMC Isolation Kit (manual separation or automated on the RoboSep™-S). For density gradient centrifugation, additional washing steps and red blood cell lysis might be required.
[...]the method of choice for PBMC isolation should be time-efficient. [...] This study showed that the number of PBMCs isolated from whole blood was about 2.44 times higher in the case of the RoboSep™-S method compared to the OptiPrep (density gradient) isolation method. This result indicates that the isolation of PBMCs by the RoboSep™-S method requires collecting a lower amount of blood from patients and saves about 50% of the time required for PBMC isolation compared to the OptiPrep® isolation method.
Go from leukapheresis pack to PBMCs in a single run with EasySep™ Direct Human PBMC Isolation Kit and RoboSep™-S. This automated cell isolation system results in fewer contaminating cells compared to density gradient centrifugation.
Figure 1. PBMC Isolation from a Full-Size Leukapheresis Pack Using the EasySep™ Direct Human PBMC Isolation Kit Automated with RoboSep™-S Results in Fewer Contaminating Cells Compared to Density Gradient Centrifugation
Mononuclear cells were isolated from single concentrated leukapheresis packs using either density gradient centrifugation with Lymphoprep™ density gradient medium (Density Gradient Centrifugation) or EasySep™ Direct Human PBMC Isolation Kit automated on the RoboSep™-S instrument (EasySep™ Direct on RoboSep™-S). Cells were counted and analyzed by flow cytometry. Compared to density gradient centrifugation, EasySep™ Direct Human PBMC Isolation Kit automated on the RoboSep™-S instrument resulted in (A) equivalent numbers of total mononuclear cells, (B) equivalent numbers of total monocytes and lymphocytes, and (C) lower numbers of residual platelets (CD41+), red blood cells (Glycophorin A+ /CD45− ), and granulocytes (CD66b+ ) (mean ± SD n=6).
See how other scientists have used RoboSep™-S to automate their PBMC isolations and help them achieve their research goals:
| Publication | Authors |
|---|---|
| Direct Haplotype-Resolved 5-Base HiFi Sequencing for Genome-Wide Profiling of Hypermethylation Outliers in a Rare Disease Cohort | Cheung W et al. |
| Cellular Stress Assay in Peripheral Blood Mononuclear Cells: Factors Influencing Its Results | Tessema B et al. |
Automating PBMC Isolations Using RoboSep-16™
The RoboSep™-16 instrument can be used to automate EasySep™ Direct PBMC isolation from up to 16 samples with minimal hands-on time. By automating all reagent and sample handling steps, and through the use of disposable pipette tips, RoboSep™-16 rapidly isolates PBMCs while minimizing the risk of sample cross-contamination.
For more information on automating PBMC isolation using RoboSep™-16, please contact techsupport@stemcell.com.
Products for Immunomagnetic PBMC Isolation
| Product | Catalog # |
|---|---|
| EasySep™ Direct Human PBMC Isolation Kit | 19654 |
| EasySep™ Direct Human PBMC Isolation Kit for RoboSep™ | 19654RF |
| RoboSep™-S | 21000 |
| RoboSep™-16 | 23000 |
Try Automated PBMC Isolation in Your Own Lab!
See for yourself how RoboSep™ can enable you to isolate PBMCs efficiently and with minimal hands-on time.
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