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Isolation of Peripheral Blood Mononuclear Cells Using Vacutainer® Cellular Preparation Tubes (CPTTM)
Alaina Puleo1, Chantia Carroll1, Holden T Maecker1
1Institute for Immunity, Transplantation, and Infection, Stanford University School of Medicine, Stanford, Palo Alto, CA, USA.
Abstract:
Peripheral blood mononuclear cell (PBMC) isolation is commonly done via density gradient centrifugation over Ficoll-Hypaque, a labor-intensive procedure that requires skilled technicians and can contribute to sample variability. Cellular Preparation Tubes (CPTs) are Vacutainer blood draw tubes that contain Ficoll-Hypaque and a gel plug that separates the Ficoll solution from the blood to be drawn. Once blood is drawn into CPTs, they can be centrifuged to separate the PBMC, then shipped (if desired) to a processing lab. The processing lab removes the PBMC from the upper compartment of the tube (above the gel plug), washes the PBMC, and can cryopreserve them using DMSO-containing media, as detailed in this protocol.
Insights
Cellular Preparation Tubes simplify peripheral blood mononuclear cell (PBMC) isolation, reducing labor and variability compared to traditional Ficoll-Hypaque centrifugation. This method enables easier PBMC separation, washing, and cryopreservation for research applications.
Area of Science:
- Immunology
- Cell Biology
- Biotechnology
Background:
- Peripheral blood mononuclear cell (PBMC) isolation traditionally uses Ficoll-Hypaque density gradient centrifugation.
- This method is labor-intensive, requires technical expertise, and can introduce sample variability.
- Cellular Preparation Tubes (CPTs) offer a streamlined alternative for blood processing.
Purpose of the Study:
- To detail a protocol for isolating and cryopreserving PBMCs using Cellular Preparation Tubes (CPTs).
- To present CPTs as a method to reduce variability and labor in PBMC isolation.
- To facilitate the shipment and processing of blood samples for PBMC recovery.
Main Methods:
- Blood is drawn into CPTs containing Ficoll-Hypaque and a gel plug.
- Centrifugation separates PBMCs within the CPT.
- PBMCs are recovered from the upper compartment, washed, and cryopreserved with DMSO-containing media.
Main Results:
- CPTs facilitate the separation of PBMCs from whole blood.
- The protocol allows for the subsequent washing and cryopreservation of isolated PBMCs.
- This method streamlines the workflow for PBMC processing and potential sample shipment.
Conclusions:
- Cellular Preparation Tubes provide an efficient and standardized method for PBMC isolation.
- This protocol simplifies PBMC preparation, suitable for various downstream research applications.
- CPTs offer a valuable tool for reducing variability and improving the accessibility of PBMC isolation.
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