|Year : 2021 | Volume
| Issue : 1 | Page : 1-5
Analysis on the composition of leukapheresis product – A comparison between MCS+® + and spectra optia® apheresis equipment
Mohandoss Murugesan1, Gayathiri K Chellaiya2, Chandran K Nair3, Sangeetha K Nayanar4
1 Department of Transfusion Medicine, Malabar Cancer Centre, Thalassery, Kerala, India
2 Department of Transfusion Medicine, Christian Medical College, Vellore, Tamil Nadu, India
3 Department of Clinical Hematology, Malabar Cancer Centre, Thalassery, Kerala, India
4 Department of Oncopathology, Malabar Cancer Centre, Thalassery, Kerala, India
|Date of Submission||10-Jun-2020|
|Date of Decision||10-Jun-2020|
|Date of Acceptance||12-Sep-2020|
|Date of Web Publication||15-Mar-2021|
Dr. Mohandoss Murugesan
Department of Transfusion Medicine, Malabar Cancer Centre, Thalassery, Kerala
Source of Support: None, Conflict of Interest: None
INTRODUCTION: Peripheral blood stem cell has become the preferred source for hematopoietic stem cells in both autologous and allogenic transplants. CD34+ cells represent a small proportion of the leukapheresis product content. The present study aims to analyze the cellular composition of leukapheresis product collected by intermittent-flow Mobile Collection System®+ (MCS+) and continuous-flow (Spectra Optia®) apheresis equipment.
METHODS: In this retrospective study, 97 leukapheresis procedures for 85 individuals mobilized only with granulocyte colony-stimulating factor were analyzed. The product samples were analyzed for CD34+ cells, red blood cells (RBCs), white blood cells, and platelet contents. The difference in overall product composition was compared between the equipment through Mann–Whitney U-test.
RESULTS: Both the equipment had similar CD34 and mononuclear cells (MNC) harvested in the product. However, Spectra Optia had statistically significantly lower product volume than MCS+ (215 mL vs. 260 mL, P = 0.04). Similarly non-CD34 composition such as RBC content per apheresis was six-fold higher (27 mL vs. 4 mL) and platelet contamination (1601 vs. 1275 × 109/L) was relatively higher with MCS+ over Spectra Optia. No relationship was observed between the CD34 concentration in the product and RBC and platelet contamination between both the equipment.
CONCLUSION: Both equipment collect adequate CD34 and MNC cells; however, Spectra Optia is preferred due to product quality in terms of less product volume with minimal RBC and platelet contamination.
Keywords: CD34, leukapheresis, MCS+® , Optia, red blood cell contamination
|How to cite this article:|
Murugesan M, Chellaiya GK, Nair CK, Nayanar SK. Analysis on the composition of leukapheresis product – A comparison between MCS+® + and spectra optia® apheresis equipment. J Appl Hematol 2021;12:1-5
|How to cite this URL:|
Murugesan M, Chellaiya GK, Nair CK, Nayanar SK. Analysis on the composition of leukapheresis product – A comparison between MCS+® + and spectra optia® apheresis equipment. J Appl Hematol [serial online] 2021 [cited 2021 Jun 23];12:1-5. Available from: https://www.jahjournal.org/text.asp?2021/12/1/1/311337
| Introduction|| |
Peripheral blood stem cell (PBSC) transplantation is performed by infusing an individual with CD34 + hematopoietic progenitor cells (HPC) and mononuclear cells (MNC) from circulating blood. This has become more common and almost replaced marrow infusion for restoring hematopoiesis in patients receiving myeloablative therapy.,,
Leukapheresis involves separation and collection of MNCs such as T cells and CD34+ HPC using apheresis equipment. The currently available apheresis equipment collect these MNCs either continuously or intermittently during the multiple cycles of collection. These equipment differ in their hardware specifications and separation technologies, but process adequate blood volume to achieve target dose for CD34 + HPC ≥2 × 106 cells/kg body weight of the recipient.
To collect MNCs, which are thought to possess a lymphocyte-like morphology, the apheresis equipment should harvest in the zone between the granulocyte layer and the platelet layer. Due to its close proximity with the platelet layer, platelet depletion can also be a concern in MNC collection. As the interface between the buffy coat zone and red blood cell (RBC) zone is narrow, RBC are being collected during the leukapheresis procedure. Collection of too many RBC may cause anemia in the individual undergoing leukapheresis.
Collection of cells such as red cells and platelets other than intended MNCs during PBSC collection is considered contaminants, and CD34 + HPC represents a small proportion of the product content. Mene´ndez et al. in their results showed that the use of different mobilization regimens not only modifies the overall number of CD34+ HPCs obtained during the leukapheresis procedures but that it also has an impact on both the absolute and the relative compositions of the leukapheresis products in different CD34+ and non-CD34 cell subsets. The present study aims to analyze the cellular composition of leukapheresis product collected between the intermittent-flow MCS+®+ (MCS+) and continuous-flow (Spectra Optia®) apheresis equipment.
| Methods|| |
It was a retrospective study conducted from January 2014 to June 2018. The leukapheresis procedure was performed in MCS+, Haemonetics (intermittent flow), and Spectra Optia, Terumo BCT (continuous flow) apheresis equipment at our center. Inclusion criteria were leukapheresis procedures for patients/donors mobilized only with granulocyte-colony-stimulating factor (G-CSF) before collection. Exclusion criteria included leukapheresis procedures performed in patients/donors mobilized with G-CSF and Plerixafor due to inadequate harvest with the first procedure or failed mobilization. These samples were excluded as Plerixafor is an independent confounding factor on CD34 yield.
In both the equipment, the precollection parameters were entered at the start of procedure (hematocrit [Hct], white blood cell [WBC], and platelet count). The target blood volume to be processed was usually set at two times the total blood volume (TBV) in both the equipment and % TBV was increased if patient had low peripheral blood CD34+ cells and WBC count on the day of collection.
Intermittent flow (MCS+®, Haemonetics) equipment
During MNC collection, the whole blood flows into the spinning bowl and plasma is drawn off until the buffy coat reaches an optical sensor. Mechanical valves divert the buffy coat into a collection bag once about 50% of the platelets have been eliminated.
Continuous flow (Spectra Optia, Terumo BCT) equipment
Collections are completely automated based on parameters entered. MNCs accumulate in a specially configured chamber within the centrifuge channel until a predetermined number of cells are reached. These cells are then automatically withdrawn into the collection bag and the tubing is rinsed with plasma.
In both the equipment, the cycle was repeated until the target blood volume was reached. The analysis was done on samples aliquoted from the leukapheresis product. The complete blood count (hemoglobin, Hct, WBC count, differential count, and platelet count) of the samples was performed using Coulter LH750 hematology analyzer, (Beckman Coulter, Brea, California, USA) in Central Lab.
The CD 34+ cell enumeration was performed on FC 500 flow cytometer using Stem Kit reagents (Beckman Coulter, Brea, California, USA) in the blood bank. The method was based on ISHAGE guidelines: four-parameter flow-cytometry method (CD45FITC/CD34PE staining, side and forward angle light scatter). Both absolute CD34 count and CD34% were recorded from the report.
The overall cellular composition of the product was analyzed. Volumes of contaminating RBCs were calculated by multiplying product volume and Hct. The product cellular composition difference between the MCS + and Spectra Optia equipment was compared through Mann–Whitney U-test. P < 0.05 was considered statistically significant. All the analyses were done using SPSS version 20 (SPSS Inc, Chicago, IL, USA).
| Results|| |
Among a total of 85 participants, 70 autologous patients and 15 allogenic donors underwent 111 leukapheresis procedures during the study period. In autologous collection, 21 patients required two procedures and two patients required a maximum of three procedures. In allogenic collection, only one donor required a second collection. Out of 111 procedures, only 97 leukapheresis (81 autologous and 16 allogenic) performed with G-CSF mobilization were analyzed. The age and diagnosis of the participants are summarized in [Table 1]. The baseline procedure parameters such as peripheral blood counts and percentage blood volumes processed were compared and were similar in both MCS+ and Spectra Optia equipment [Table 2].
|Table 1: Demographic details of patients and donors underwent leukapheresis procedure at a tertiary cancer center in South India (n=85)|
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|Table 2: Baseline characteristics of patients and donors who underwent leukapheresis using two apheresis equipment at a tertiary care cancer center in South India|
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The product volume in procedures performed with MCS + and Spectra Optia equipment was 260 mL versus 215 mL (P = 0.04). Similarly, RBC contamination in terms of milliliter per apheresis procedure was much lower with that of Spectra Optia equipment (4 mL vs. 27 mL; P < 0.01) [Table 3]. There was no difference in WBC subpopulation in the products collected between two equipment [Table 3]. Both the equipment products had no statistically significant difference in CD34 yield (P = 0.90) and MNC yield (P = 0.48) in terms of per kg body weight of the recipient available for the transplant. The total nucleated cell dose was significantly higher with the products of MCS+ equipment as these products contained high WBC content (10.3 vs. 6.9 × 108/kg body weight, P < 0.01).
|Table 3: Leukapheresis product comparison between two apheresis equipment from a tertiary care cancer center in South India|
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[Table 4] shows that the leukapheresis products were categorized into high and low concentration based on the median CD34%. The MCS + equipment had no differences in RBC and platelet contamination over product CD34 cell concentration. However, Spectra Optia had relatively higher platelet contamination when the product CD34 concentration increased.
|Table 4: Relationship of CD34 concentration in leukapheresis product versus red cell content and platelet contamination between two apheresis equipment|
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| Discussion|| |
There are several studies that had focused on apheresis equipment performance on PBSC collection using parameters such as collection efficiency. However, the present study compared the difference in the composition of leukapheresis product collected using Spectra Optia, (continuous flow) and MCS+ (intermittent flow) apheresis equipment.
The study observed no difference in CD34+ stem cells collected in product in terms of faster hematopoietic reconstitution between the two apheresis equipment. Nevertheless, there are differences between these two equipment and the products they collect. Most significant differences were observed in product volume and RBC contamination [Table 3].
Compared to marrow harvest, CD34+ stem cells from leukaphereis product represent small proportion of immature peripheral blood progenitor cells and major population of lineage committed precursors responsible for both long and short term, respectively. In the present study, the products from both the Spectra Optia and MCS+ equipment had similar levels of median CD34+ cell counts (1185 vs. 880, P = 0.42), respectively. The non-CD34+ cell composition in the product may vary based on instrument collection methodologies and donor characteristics.
The volumes of contaminating RBC in products were higher with MCS+ equipment, and there was around six-fold higher volume of RBC contamination per apheresis with MCS+ than Spectra Optia. In allogenic transplantation, RBC depletion is the most commonly performed procedure on the product before infusion, when there is ABO incompatibility. The maximum allowable RBC volume varies between centers; it is usually between 10 and 40 mL. The maximum volume of incompatible RBCs in ABO incompatible allogenic transplant that can be safely infused is not clearly defined and the usual limit is 20–30 mL or 0.2–0.4 mL/kg. It has been demonstrated that when <15 mL of residual RBCs is infused, no clinically significant acute hemolysis occurs. Hence, the products from Spectra Optia can be used without manipulation in ABO incompatibility setting.
Spectra Optia had statistically significantly low product volume than MCS+, thereby facilitating minimum storage space during cryopreservation (P = 0.04). Another advantage is lower volume of cryopreserved products reduces the amount of Dimethyl sulfoxide (DMSO) infused into the recipient. Studies had shown that red cells in product interfere with tumor purging technique that relies on complement fixation or affect positive selection of cells in graft engineering of stem cells.
The main difference in such RBC contamination was that Spectra Optia utilizes automated interface management system, an optical detection system composed of hardware and software, that interprets the interface position in tubing set connector during the procedures. However, in MCS+, the parameters Vol into RBC/collec and Vol into RBC/recirc determine the volume of RBC to be collected after the line sensor detects the RBC detection. In general, more buffy coat gets collected with increasing collect volume, thereby granulocyte and red cell content of the product increases.
Cytopenias, typically thrombocytopenia, are well-recognized transient effects of leukapheresis. Decrease in platelet count was observed in 30%–50% after first apheresis collection., There are two reasons for the minimized platelet content of the leukapheresis products. Platelet content in the product is monitored to prevent thrombocytopenia in patients and high platelet count in the product may cause clumping. The study observed that the number of platelets contaminated in the product was lesser with Spectra Optia over MCS+ (1275 vs. 1601, P = 0.03). A literature review showed that automated platform for PBSC collection tends to have lower platelet collection over the manual platform.
Granulocytes in product release peroxidases and other toxic compounds that could reduce stem cell viability on lyse and thaw during cryopreservation. The products in the present study had no difference in their granulocyte concentration between the equipment (P = 0.64).
Being a retrospective study, there are limitations with the record review. Identification of WBC subpopulation such as NK cells and progenitor cells was not practiced at the center. Similar studies with prospective nature on WBC subpopulation identifying types of HPC can be carried out to study the composition of these populations in the product.
| Conclusion|| |
The ideal apheresis equipment for leukapheresis procedure should have high selectivity and efficiency for MNC collection. Both MCS+ and Spectra Optia equipment collect sufficient CD34+ cells that are adequate for PBSC transplant. The automated program in Spectra Optia is preferred over the manual program of MCS+, preferably due to the quality of the product in terms of contamination of RBC, platelets, and low product volume.
The authors thank Mrs. Maya and Mr. Riyas, and the staffs of Blood Bank, Malabar Cancer Centre (MCC), for their assistance in data collection and analysis. The authors also thank the patients registered in MCC whose undisclosed data helped to carry out this research.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
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[Table 1], [Table 2], [Table 3], [Table 4]