|Year : 2017 | Volume
| Issue : 2 | Page : 102-108
Comparison of “Amicus and COBE Spectra” for autologous peripheral blood stem cell harvest: An Indian experience
Rasika Dhawan Setia1, Satyam Arora2, Anil Handoo3, Dharma Choudhary4, Sajeev Kumar Sharma4, Tina Dadu3, Divya Doval4, Meenu Kapoor1, Shalu Bajaj1, Virendra Bachchas3
1 Department of Transfusion Medicine and Hematology, BLK Super Speciality Hospital, New Delhi, India
2 Department of Transfusion Medicine, Super Speciality Pediatric Hospital and Postgraduate Teaching Institute, Noida, Uttar Pradesh, India
3 Department of Hematolgy, BLK Super Speciality Hospital, New Delhi, India
4 Department of Hemato-Oncology and Bone Marrow Transplant, BLK Super Speciality Hospital, New Delhi, India
|Date of Web Publication||11-Sep-2017|
Department of Transfusion Medicine, Super Speciality Pediatric Hospital and Postgraduate Teaching Institute, Noida, Uttar Pradesh
Source of Support: None, Conflict of Interest: None
Background: Adequate collection of peripheral stem cells from the patients depends on the disease condition, efficient mobilization and the equipment used for PBSC harvest. COBE Spectra has been the major platform for collecting these PBSC for more than 2 decades. Now with introduction of PBSC harvest option with Amicus cell separators (which was used primarily for plateletpheresis) in India, it is worth while comparing both the platforms for stem cells collections. Materials and Methods: Our study is a retrospective analysis of autologous PBSC harvest procedures done at our centre. The study included the data of autologous PBSC collections from January 2015 to June 2016. Total 61 patients underwent 85 autologous PBSC harvests for both haematological and non haematological indications. Results: Out 61 patients, 40 patients collected their target number of cell in a single harvest, 18 patient required dual harvests and 3 patients required three consecutive days of harvest. Pre-Apheresis WBC, platelets and CD45/CD34 cell counts were comparable. COBE Spectra collects significantly higher product volume and higher number of platelets in the apheresis product. Whereas WBC counts of the product, total CD45/CD34 cell dose and collection efficiency (CE2) and collection ratio (CR) were comparable on both the platforms. Conclusions: Amicus took more time to harvest the anticipated number of cells in the graft as well as COBE Spectra resulted in higher platelets loss during the process of cell collection. Our analysis is first of its kind from Indian subcontinent and indicates that with gradual phasing out of COBE spectra from the market Amicus offers a comparative platform for PBSC harvest.
Keywords: Amicus, autologous stem cell transplant, COBE Spectra, peripheral blood stem cells, stem cell harvest
|How to cite this article:|
Setia RD, Arora S, Handoo A, Choudhary D, Sharma SK, Dadu T, Doval D, Kapoor M, Bajaj S, Bachchas V. Comparison of “Amicus and COBE Spectra” for autologous peripheral blood stem cell harvest: An Indian experience. Glob J Transfus Med 2017;2:102-8
|How to cite this URL:|
Setia RD, Arora S, Handoo A, Choudhary D, Sharma SK, Dadu T, Doval D, Kapoor M, Bajaj S, Bachchas V. Comparison of “Amicus and COBE Spectra” for autologous peripheral blood stem cell harvest: An Indian experience. Glob J Transfus Med [serial online] 2017 [cited 2018 Apr 26];2:102-8. Available from: http://www.gjtmonline.com/text.asp?2017/2/2/102/214285
| Introduction|| |
Autologous hematopoietic stem cell transplantation (AHSCT) has been used to treat various hematological and nonhematological malignancies. At present, majorly AHSCT is performed with stem cells harvested by peripheral blood stem cells (PBSCs) from growth factors or chemotherapy-mobilized patients. Minimum dose required for a successful transplant is 2 × 106 CD34+ cells/Kg of the patient whereas higher doses have also been associated with better recovery and early engraftments.
Adequate collection of these cells from the patients depends on the disease condition, efficient mobilization, and the equipment used for PBSC harvest. COBE Spectra has been the major platform for collecting these PBSCs for more than two decades. Now with the introduction of PBSC harvest option with Amicus cell separators (which was used primarily for plateletpheresis) in India, it is worthwhile comparing both the platforms for stem cell collections. Our study is a retrospective analysis of autologous PBSC harvest procedures done at our center. We have analyzed and compared the factors affecting autologous harvest on both platforms and the cellular variation of the autologous PBSC products harvested.
| Materials and Methods|| |
Demography of the patients
The study included the data of autologous PBSC collections at our center from January 2015 to June 2016. The indications for AHSCT were both hematological and nonhematological conditions. All the patients were evaluated by the bone marrow transplant physician and apheresis physician for adequate suitability for PBSC harvests as well as necessary laboratory tests were done before initiating the mobilization regimen. All the patients had given an informed consent in accordance with the Declaration of Helsinki.
For mobilization of PBSC, we used human granulocyte-colony-stimulating factors (GCSFs; 10 μg/kg/day single doses) administered subcutaneously (SC) starting 5 days before leukapheresis. A circulating CD34+ cells level greater than 10 × 106/L or total white blood cells (WBCs) count >20 × 109/L were used as the main criterion for adequate mobilization and beginning of leukapheresis. In case of poor mobilization, Plerixafor (0.24 mg/kg SC) was added to the mobilization regimen on day 5 or 6 with GCSF. In cases of inadequate collection of CD34+ cell dose on 1st day of collection, another session of leukapheresis was done on the next day till the target dose was achieved.
Peripheral blood stem cell collections (leukapheresis)
The apheresis platforms used were COBE Spectra and Amicus. The patients were randomly assigned the machines. Amicus equipment is based on a continuous flow method controlled by computer software. Amicus device consists of a single chamber designed to collect PBSC, but the collection occurs in phases. COBE Spectra device uses “optimization” algorithm for maximizing CD34+ cells recovery as well as the algorithm is based on pre-Mononuclear cells count and inlet pump rate guiding flow rates. COBE Spectra is operator dependent for adjustment based on the color scale; hence, it is a semi-automated device lacking optical sensors for collection monitoring. The apheresis procedure was performed according to the respective manufacturer's standard operating procedure. Apheresis procedure was stopped when the processing volume reached 2–3 times of total blood volume (TBV).
Blood cell counts (both preharvest peripheral blood (PB) and of the graft) were done using LH750 Beckman Coulter (Florida, Miami, USA). CD45+ and CD34+ cell count was done using BD FACSCanto-II Flow Cytometer. Enumeration of CD34+ cells was done by flow cytometry as described by International Society of Hematology and Graft Engineering guidelines.
Collection efficiency (CE/CE2) was calculated to compare the effectiveness of stem cell extraction with different systems. CE2 was calculated using following formula:
Total CD34+ cells in the product were calculated by multiplying CD34+ (/μL) in the product and the volume of the product (mL). PB CD34+ cells were the concentration of CD34+ cell present in the PB before the apheresis. Apheresis volume processed was the volume of whole blood processed by the apheresis machine subtracting acid citrate dextrose (ACD) used.
Collection ratio (CR) was also calculated, as another parameter to evaluate the efficiency of the stem cell collection. CR was calculated as the number of target cells in the product bag over the concentration of target cells in the circulation.
CR = (CD 34 Positive cells (/kg) of product]/PB CD 34 Positive cells (μL)
Platelet CR (Plt CR %) was another parameter which was evaluated to study the platelet loss in the harvested product among both the platforms. It was calculated similarly as CE2. Dose of the PBSC product is number of CD34 positive cells collected per liter per kilogram of the patient.
The data were expressed as the mean ± standard deviation (SD) and median (range). Data analysis was done using Excel software (2007) by Microsoft. Unpaired t-test was used for intergroup data analysis and P< 0.05 was considered statistically significant difference between the groups. Linear regression analysis was used to study the association of donor variables such as preharvest CD34+ cells with total CD34+ cells collected and the collection efficiency of the both the cell separators.
| Results|| |
Demography of the patients
A total of 61 patients underwent 85 autologous PBSC harvests for both hematological and nonhematological indications during the period of analysis. [Table 1] discusses the various indications and disease condition of the patients undergoing autologous transplants. Out of 61 patients, 40 patients collected their target number of cell in a single harvest, 18 patient required dual harvests, and 3 patients required 3 consecutive days of harvest [Table 2]. Thirty-eight patients out of 61 were male and 23 were females [Table 3]. Mean ± SD and median of age of patients analyzed were 41.7 ± 18.3 years and 47 years with a range from 3 years to 67 years. For comparison purpose, we have only analyzed the first harvest of our patients (n = 61).
|Table 2: Number of apheresis sessions required by the patients based on type of cell separator and mobilization regimen used|
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|Table 3: Comparison between COBE Spectra and Amicus platforms for autologous stem cells harvests|
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Preapheresis and apheresis procedural characteristics
[Table 3] discusses the PB counts before the first harvest of all the patients before their first apheresis harvests. Preapheresis WBC, platelets, and CD45/CD34 cell counts were comparable, and there was no statistically significant difference among patients undergoing apheresis by both the equipment. TBV processed and total ACD infused also did not show any statistical difference whereas patients undergoing harvest on Amicus took more time to collect the anticipated dose when compared with COBE Spectra (P < 0.05).
Apheresis product analysis
Apheresis product from both the cell separators has been discussed in [Table 3]. COBE Spectra collects significantly higher product volume and a higher number of platelets in the apheresis product, whereas WBC counts of the product, total CD45/CD34 cell dose, and collection efficiency (CE2) and CR were comparable on both the platforms.
Factors affecting autologous peripheral blood stem cells collection
Preapheresis CD45/CD34 cell counts
[Table 4] discusses the effect of PB preapheresis CD45/CD34 cell counts on PBSC harvest of both the platforms. Patients harvested with COBE Spectra (total n = 40) having higher (>40 × 106/L) preapheresis PB CD45/CD34 cell counts (n = 20) showed significantly higher preapheresis PB WBC, platelets, and CD45/CD34 cells counts and consecutively collected significantly higher WBC, platelets and cell dose in the apheresis products compared to group (n = 20) with lower (=<40 × 106/L) PB CD45/CD34 cell counts, whereas collection efficiency (CE2) and CR both were higher in the group with lower PB CD45/CD34 cell counts.
|Table 4: Comparison between COBE Spectra and Amicus platforms based on pre-CD34+ cell counts for autologous stem cells harvests|
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Patients harvested with Amicus (total n = 21) having higher (>40 × 106/L) PB CD45/CD34 cell counts (n = 11) showed significantly higher CD45/CD34 cells counts and consecutively collecting significantly higher cell dose in the apheresis products compared to the group with lower (≤40 × 106/L) PB CD45/CD34 cell counts (n = 10), whereas collection efficiency (CE2) and CR both were higher in the group with lower (≤40 × 106/L) PB CD45/CD34 cell counts when harvested with Amicus as with COBE Spectra.
In over all cases (both on Amicus and COBE) with preapheresis PB CD45/CD34 cell counts ≤40 × 106/L (n = 31), patients harvested on Amicus (n = 11) showed significantly higher preapheresis platelet counts and took more time for the harvests but platelets counts were still significantly higher in product collected by COBE Spectra (n = 20).
In over all cases (both on Amicus and COBE) with preapheresis PB CD45/CD34 cell counts > 40 × 106/L (n = 30), patients harvested on Amicus (n = 10) took significantly more time for the harvests but platelet counts and apheresis product volume were still significantly higher in product collected by COBE Spectra (n = 20).
[Table 5] discusses the effect of mobilization regimens on PBSC harvest of both the platforms. Patients harvested with COBE Spectra (total n = 39) mobilized with GCSF with Plerixafor (n = 16) showed significantly higher preapheresis PB WBC, CD45/CD34 cell counts and consecutively collected significantly higher WBC, lymphocytes, and cell dose in the apheresis products, whereas the group of patients with GCSF only mobilization (n = 23) processed significantly higher blood volume.
|Table 5: Comparison between COBE Spectra and Amicus platforms based on mobilization regimen for autologous stem cells harvests|
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Patients harvested with Amicus (total n = 21) mobilized with GCSF and Plerixafor (n = 07) showed significantly higher cell dose collection in the apheresis products. Whereas the group of patients with GCSF only mobilization (n = 14) took more time to collected the cell dose as well as had significantly better collection efficiency and CR.
In over all cases with GCSF mobilization (n = 37), patients harvested on Amicus (n = 14) showed significantly more time for harvest and collected higher lymphocytes in the apheresis product, whereas platelets counts were still significantly higher in product collected by COBE Spectra (n = 23).
In over all cases with GCSF with Plerixafor mobilization (n = 23), patients harvested on Amicus (n = 07) showed significantly more time for harvest, whereas products collected by COBE Spectra (n = 16) showed significantly higher product volume, product platelets count, collection efficiency, and CR.
Inadequate first collections
Twenty-one patients out of 61 (34.4%) required an additional session of PBSC harvest on the subsequent days. Details of their diagnosis are discussed in [Table 1]. Eighteen (13:5, male:female) out of 21 required one additional session (total two), with their age ranging from 3 to 63 years. Three patients (1:2, male:female) required total three sessions to complete their cell collection with their age ranging from 9 to 48 years. Mean dose of CD45/CD34+ cells collected on first harvest for patients requiring dual session (n = 18) was 2 × 106/Kg/L and patients requiring triple session (n = 3) were 0.46 × 106/Kg/L as compared to overall (n = 61) dose 4.8 × 106/Kg/L collected.
| Discussion|| |
Our analysis compared COBE Spectra and Amicus apheresis platforms for autologous PBSC harvests. The demographic details as well as baseline PB counts preapheresis of patients in both the groups were comparable [Table 3]. The comparison showed that both machines are efficient in harvesting peripheral stem cells in autologous transplant settings. There was no statistical difference in the amount of whole blood processed and ACD exposure on both the platforms whereas Amicus took more time to complete the collection process. One of the similar comparative studies  showed significantly higher product volume collected by Amicus when compared to COBE Spectra, in contrast to our results [Table 3], was due to significantly higher blood volume processed by Amicus in that study.
As previously reported , our study also showed COBE Spectra resulting in higher apheresis-induced thrombocytopenia during PBSC harvests, as compared to Amicus. This was due to the difference in the process of cell separation and harvest by both of the platforms. Cellular lymphomononuclear population of the apheresis product by both platforms was comparable as well as the CD45/CD34 collected dose. CE2 prediction (Benchmark “CE2;” first described by Pierelli et al.) is based on the fact that CD34+ cells released from the bone marrow at a constant rate and CE2 is the efficiency of any apheresis platform to capture that stem cell in the harvest product. The release of CD34+ cell in the circulation is not constant as well as varies with regard to the mobilization regimen and the underlying condition of the marrow. Hence, calculating and validating CE2 for each platform is necessary for its application for accurate prediction. Our analysis showed comparable CE2 on both the platforms [Table 3]; (49.9 ± 26.1% COBE vs. 55.2 ± 41.5% Amicus).
We analyzed few important factors associated with autologous PBSC harvest. One of the most important factors is pre-CD34+ cell concentration before initiating apheresis session [Table 4]. Higher preharvest WBC counts, platelet counts, and CD45/CD34+ cell counts (>40 × 106/L/) were associated with significantly higher cell counts and dose in the grafts harvested whereas a lower precount (≤40 × 106/L) was associated with better CE2 and CR, irrespective of the platform used. COBE Spectra harvested higher platelets in the graft irrespective of higher preplatelet counts of patients using Amicus as cell separator.
Another important factor analyzed was the mobilization regimen. Mobilization regimen including GCSF with Plerixafor was associated with better mobilization (higher peripheral cell counts) preharvest, hence resulting in higher cell counts in the graft when compared to GCSF only regimen. GCSF only regimen took significantly more time in completing the harvest as due to low precounts more blood volume (not statistically significant) was targeted. Irrespective of the regimen used Amicus took more time to complete the harvests when compared to COBE Spectra. This may be due to the collection and harvest of stem cells in a cycled format in Amicus.
A total of five patients (8.1%) out of 61 reported with adverse reactions to PBSC harvests, 3 on COBE Spectra and 2 on Amicus. Four out of five patients presented with chills, rigors, and hypotension (signs and symptoms of calcium depletion) and one patient (on COBE) presented with a significant reduction of platelet counts (<20 × 103/μL), hence requiring transfusion of platelets postprocedure. All the harvests were done with central line as the mode of venous access. The main limitation of our analysis is the retrospective nature of the study as well as inability to calculate other parameters such as CE1 and platelets attrition (%) for a complete comparison.
| Conclusion|| |
Our study suggests that both Amicus and COBE Spectra offer comparable results from autologous PBSC collections in terms of collection efficiency, CR, absolute CD34+/CD3+ cells, and stem cell dose. Amicus took more time to harvest the anticipated number of cells in the graft as well as COBE Spectra resulted in higher platelets loss during the process of cell collection. Both precell counts in PB and mobilization regimen significantly affected the PBSC harvest on both the platforms. Our analysis is of its kind (COBE vs. Amicus) from Indian subcontinent and indicates that with gradual phasing out of COBE Spectra from the market Amicus offers a comparative platform for PBSC harvest.
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], [Table 5]