Establishment of a cell processing laboratory to support hematopoietic stem cell transplantation and chimeric antigen receptor (CAR)-T cell therapy

Role of flow cytometry in evaluation of the cellular therapy products used in haematopoietic stem cell transplantation

“…Lymphocytes for DLI could be collected during allo‐HSC collection procedure or from unstimulated PB, and cryopreserved as a simple and effective therapeutic option for patients in case of disease relapse 40,41 . The purpose of DLI is to enhance donor T‐cells potency against leukaemic cells (graft‐vs.‐tumour effect), that is, to treat disease relapse and improve immune recovery 42 …”

“…As with CD34+ cells collection for HSC transplantation, obtaining a sufficient concentration of T‐cells is a critical part of the collection process. It is therefore important to perform quality control of leukapheresis products using anti‐CD3 mAb, because quality of the final product depends on the quality of the starting material used in CAR‐T cells manufacturing 42,52 . After the manufacturing process, a sample of the CAR‐T cell product is taken for quality control, which includes, among other things, phenotyping, viability and purity of effector cell population 42 …”

“…40,41 The purpose of DLI is to enhance donor T-cells potency against leukaemic cells (graft-vs.tumour effect), that is, to treat disease relapse and improve immune recovery. 42 In the fresh apheresis products which will be cryopreserved for DLI, CD3+ cells are determined on flow cytometer. Since there are only a few reports on how this cell population tolerates the cryopreservation and thaw process, and because of difference among laboratories in cryopreservation protocols, each centre should determine the viability of CD3+ cells in the thawed sample before infusion.…”

“…It is therefore important to perform quality control of leukapheresis products using anti-CD3 mAb, because quality of the final product depends on the quality of the starting material used in CAR-T cells manufacturing. 42,52 After the manufacturing process, a sample of the CAR-T cell product is taken for quality control, which includes, among other things, phenotyping, viability and purity of effector cell population. 42 In addition to the determination of the quality of the leukapheresis product, flow cytometry is used in the monitoring of expansion and persistence of the therapeutic cells after the infusion and in the evaluation of treatment response in patients who received graft with CAR-T cells.…”