Evaluation of ‘out-of-specification’ CliniMACS CD34-selection procedures of hematopoietic progenitor cell-apheresis products
Introduction
Transplantation of positively selected CD34+ hematopoietic progenitor cells (HPC) is part of an established treatment modality for adult and pediatric patients with hematologic malignancies. Immunomagnetic selection of CD34+ HPC provides a graft profoundly depleted of T, B and natural killer (NK) cells, which reduces the incidence of both acute and chronic graft-versus-host disease (GvHD) after allogeneic stem cell transplantation (SCT) 1., 2., 3.. In autologous SCT, selection of CD34+ cells is used both to reduce the tumor cell contamination of the graft in CD34− malignancies, which may decrease relapse risk [4], and to deplete T cells in grafts used in autoimmune diseases [5, 6].
Several studies have shown that the CliniMACS device is a reliable system for clinical-scale selection of large amounts of CD34+ cells, yielding high purities and good recoveries 7., 8., 9., 10., 11.. The capacity of the disposable tubing sets and the separation capacity of 1 vial of CD34 reagent determine the number of selection procedures required and the total cost of the processing procedure. According to the manufacturer's current specifications, the maximum capacity of a normal-scale tubing set is 600 × 106 CD34+ cells and 60 × 109 total nucleated cells, whereas that of the large-scale tubing set is 1200 × 106 CD34+ cells and 120 × 109 total nucleated cells. The specified separation capacity of 1 vial of CliniMACS CD34 reagent is 600 × 106 CD34+ cells out of 60 × 109 total nucleated cells. Single or pooled HPC-apheresis (HPC-A) products from the same donor frequently exceed one of the limits of the tubing set and/or 1 vial of CD34 reagent. Therefore, many stem cell-processing laboratories in Europe regularly overload tubing sets and/or use less CD34 reagent than recommended to reduce the number of selection procedures, processing time and total cost.
In this study, we retrospectively evaluated the performance, in terms of CD34+ cell recoveries and purities and T-cell depletion efficiencies, of the CliniMACS system in a large number of ‘out-of-specification’ CD34-selection procedures performed in eight stem cell-processing laboratories.
Section snippets
Selection of CD34+ cells
Six-hundred and forty-three HPC-A products were collected from healthy donors (n=609) and patients with solid tumors or autoimmune disease (n = 34). Almost all the HPC-A products were processed within 24 h of harvest. Some HPC-A products harvested from unrelated healthy donors abroad were processed between 24 and 48 h after harvest. Positive selection of CD34+ cells was performed using the CliniMACS® device (Miltenyi Biotec, Bergisch Gladbach, Germany), according to the manufacturer's standard
CD34+ cell recovery of overloaded normal- and large-scale tubing sets
In the eight participating stem cell-processing laboratories, 643 CliniMACS CD34-selection procedures were performed on HPC-A products. Two-hundred and fifty-three of these procedures were outside the limits of the tubing set capacity specified by the manufacturer. This allowed us to evaluate the effect of overloading both types of tubing sets with either excess amounts of total nucleated cells or excess amounts of CD34+ cells or both. Therefore we divided the CD34-selection procedures based on
Discussion
In this study, we evaluated a large number of out-of-specification CliniMACS CD34 selection procedures of HPC-A products for CD34+ cell recovery and purity, and compared the results with those of within-specification procedures. Our study supports three main conclusions. First, overloading normal- or large-scale tubing sets with excess numbers of nucleated cells does not hamper the recovery and purity of CD34+ cells. Secondly, overloading normal- or large-scale tubing sets with excess numbers
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Characterization of a genomic region 8 kb downstream of GFI1B associated with myeloproliferative neoplasms
2021, Biochimica et Biophysica Acta - Molecular Basis of DiseaseCitation Excerpt :This study was approved by the Medical Ethics Committee of the RadboudUMC and conducted in accordance to the Declaration of Helsinki. Positive selection of CD34+ cells was performed using the CliniMACS device (Miltenyi Biotec, Bergisch Gladbach, Germany) according to manufacturer's procedure [see [23]]. BFU-E and CFU-GM were evaluated in a methylcellulose-based medium (Methocult H4534; Stem Cell Technologies, Vancouver, Canada).
Cell processing for haplo-identical hematopoietic stem cell transplantation: Automated washing and immunomagnetic-positive selection
2012, CytotherapyCitation Excerpt :Immunomagnetic cell selection (ICS) of CD34+ cells, collected by leukapheresis (hematopoietic progenitor cell apheresis; HPC-A), is being used increasingly in autologous and allogeneic transplant settings. In the first case, the ICS is adopted in order to reduce neoplastic contamination of CD34– cells (1,2), while in the second case, and in particular in haplo-identical transplantation, the aim is to reduce T-cell quantity and incidence of graft-versus-host disease (GvHD) (3,4). Moreover, the T- and B-cell depletion reduces the risk of developing Epstein–Barr virus (EBV)-induced lymphoma.
Characteristics of CliniMACS<sup>®</sup> System CD34-Enriched T Cell-Depleted Grafts in a Multicenter Trial for Acute Myeloid Leukemia-Blood and Marrow Transplant Clinical Trials Network (BMT CTN) Protocol 0303
2012, Biology of Blood and Marrow TransplantationCitation Excerpt :Two of the 3 products that exceeded the loading limit by more than 10% had <50% recovery, indicating that this upper limit for CD34+ cells should not be exceeded. Our observations are in agreement with a recently published analysis of a large number of products processed at multiple centers showing that TNC in excess of that recommended by the manufacturer does not affect device performance, whereas exceeding the recommended CD34+ cell limit reduces CD34+ cell recovery [10]. Even though differences among centers were seen in the statistical analysis, all centers were able to process final graft products that easily met the criteria of the study.
Positive immunomagnetic CD34<sup>+</sup> cell selection in haplo-identical transplants in β-thalassemia patients: Removal of platelets using an automated system
2010, CytotherapyCitation Excerpt :Immunomagnetic CD34+ stem cell selection (ICS) harvested by leukapheresis (LKF) is used in autologous transplants in order to reduce contamination of negative neoplastic CD34 cells (1,2), while in allogeneic transplants, in particular haplo-identical transplants, the goal of the selection is to reduce the quantity of T cells in order to diminish the occurrence of graft-versus-host disease (GvHD) and eventually to modulate graft-versus-leukemia (GvL) (3,4).