Peripheral Blood Mononuclear Cells (PBMCs)

You can find our PBMCs Protocols by clicking here.

We also have the ability to follow our clients’ custom processing protocols and help them develop new processing protocols based on their needs.

Our standard collection tube for PBMCs is a Sodium Heparin Tube; however, prospective collections can utilize any commercially available blood collection tube.

We recommend shipping PBMCs on dry ice or, if possible, in liquid nitrogen. PBMCs should be stored in liquid nitrogen immediately upon receipt.

Red blood cell lysis is not performed.

Cell counts and viability are determined using a Nexcelom Cellometer® with acridine orange and propidium iodide to identify live and dead nucleated cells, respectively. Unlike dissociated tissue, which is prone to cellular debris, PBMCs can be counted using trypan blue exclusion. If flow cytometry is used to determine cell count and viability, it is recommended that proper FSC thresholds be established to remove platelets, which are present in cryopreserved PBMCs.

Inventory PBMCs are collected using sodium heparin as the anticoagulant. Alternative anticoagulants, such as EDTA and sodium citrate, are available upon request.

Diseased PBMC and BMMC samples are occasionally collected at a different date from the initial sample utilized to establish blast percentages. Therefore, DLS is unable to guarantee blast percentages in diseased samples. However, for AML and multiple myeloma BMMCs, DLS performs flow cytometry following cryopreservation to characterize blast percentages, and these results are available to aid in sample selection.

Following at least 24 hours in liquid nitrogen, one vial is removed from storage and quickly thawed in a 37°C water bath until only a small frozen crystal remains. Vials are transferred into a biosafety cabinet and diluted with an appropriate volume of DMEM/F12 + 10% FBS in a 15 mL conical tube to ensure linearity with the Nexcelom Cellometer. 20 µL of the cell suspension is mixed with 20 µL of acridine orange/propidium iodide and counted on the Nexcelom Cellometer to determine cell counts and viability. All cell counts are performed prior to any pelleting and washing of the samples. These counts are estimations of the total live cell yield.

PBMCs are shipped on dry ice. Upon receipt, they should be used immediately or placed in liquid nitrogen vapor phase for long term storage.

Peripheral blood mononuclear cells (PBMCs) are white blood cells with round nuclei—primarily lymphocytes (T cells, B cells, NK cells), monocytes, and dendritic cells—routinely isolated for immunology and cell therapy research. They are commonly prepared as a first step prior to isolating specific immune subsets or performing functional assays.

Discovery PBMCs are collected under controlled SOPs (whole blood or leukopaks), isolated by density gradient methods (e.g., Ficoll-based), and cryopreserved the same day to minimize granulocyte carryover and preserve function. Each lot undergoes flow-cytometric QC for viability, purity, and major subset frequencies prior to release.

Rapidly thaw vials at 37°C, dilute promptly in recovery medium, and spin to remove cryoprotectant. Seed or rest cells per your assay. For accurate live-cell counts—especially with PBMCs—use AO/PI staining with an automated counter to enumerate nucleated cells and avoid trypan-blue under/over-calling. A short video SOP is available on request.

Yes. Discovery can provide HLA Class I/II-typed, viral-negative PBMCs from healthy or disease-state donors, and we can supply matched plasma/serum when required. Ask us about pre-screened responsiveness (e.g., PHA/CEF) and demographic/clinical metadata to align donors with your study design.