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.

At Discovery Life Sciences, peripheral blood mononuclear cells (PBMCs) notates cells isolated from whole blood, while mononuclear cells (MNCs) represent cells isolated from apheresis material. During the apheresis process, granulocytes, red blood cells, and platelets are reduced in the specimen, and are further removed during the processing. Therefore, MNCs will have drastically reduced levels of platelets compared to whole blood 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 15ml conical tube to ensure linearity with the Nexcelom Cellometer.  20µl of the cell suspension is mixed with 20µl 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.