A Single Center 25-year Experience in Autologous Peripheral Blood Stem Cell Collection: A Focus on the Collection Efficiency

Background:Harvest of hematopoietic progenitor cells via leukapheresis is being used increasingly for autologous transplantation. Adequate yield of cells per kilogram body weight of recipient is required for a successful engraftment. Collection efficiency(CE)is a useful parameter to assess quality of peripheral blood stem cell (PBSC) collection program. In this study, we report a 25-year experience in a tertiary care Hospital in Italy.Patients and Methods:1,026 consecutives autologous PBSC collection procedure, performed in 763 patients, from January 1996 to December 2020 were retrospectively considered.Data regarding patients, Blood Cells Separators (BCS) , apheresisprocedures and PBSC products were collected in our database. In these 25 years different BCSwere adopted in our Apheresis Unit (AU). In the first period (1996-1999) we used Fresenius Com.Tec, in the central period (2000-2013) we used Cobe Spectra and inthe last period (2014-2020) Spectra Optia. Results:As regards the evaluation of patients before leukapheresis, the most significant data was the increasing number of CD34+ cells. Considering the PBSC collection procedure, there was a progressive increase in the processed blood volume with a shorter apheresis duration. Data related to the PBSC collection demonstrated an increasing CD34+ cell yield and efficiency a raise in CE that was 43% using Fresenius COM-TEC BCS, 49% using Cobe Spectra BCS and 53% using Spectra Optia BCS. . Conclusions:These results were observed considering a 25-year period, thus a great number of factors likely contributing to the observed results, including technological improvement of the instrumentation for leukapheresis,increased experience of the team operating in the Apheresis Unit, improved mobilization protocols, better criteria for patients’ selection. Focusing our attention on CE we observed quite satisfactory results with a median which rose from 43% to 53% with an increase of 10% in the observation period.

Blood coagulation function before and after peripheral blood stem cell mobilization and collection / 中国输血杂志

【Objective】 To study the changes of blood coagulation function of donors before and after peripheral blood stem cell(PBSC)mobilization and collection, so as to evaluate the safety of the current scheme. 【Methods】 30 donors who received PBSC mobilization and collection in Zhujiang Hospital from October 2018 to October 2020 were enrolled. After mobilization by G-CSF, the correlation between coagulation function, blood routine indexes and TEG indexes of donors was analyzed, and the influence of PBSC mobilization and collection on coagulation function of donors was evaluated. 【Results】 The TEG indexes R(min), K(min), α(°), MA(mm) and CI before and after PBSC collection were 6.12±1.18 vs 7.25±2.16, 1.98±0.41 vs 2.45±0.64, 62.82±4.98 vs 57.3±6.67, 60.93±3.26 vs 55.37±4.41, and -0.31±1.40 vs -2.32±2.18, respectively(P<0.05), suggesting that there was no risk of hypercoagulability after PBSC mobilization and collection. The peak values of WBC (×109/L), Plt (×109/L) and Hb (g/L) were 62.02, 357 and 162, respectively, which indicated that the blood routine indexes after PBSC mobilization and collection were in the safe range. After PBSC collection, the CI value of 26.7% (8/30) donors was less than -3, showing hypocoagulability. 【Conclusion】 The current mobilization and collection scheme of PBSC has little effect on the coagulation function. Most of the donors had no risk of hypercoagulability, but a few showed a trend of hypocoagulability after PBSC collection.

Factors Influencing Peripheral Blood Stem Cell Collection / 종양간호학회지

PURPOSE: Peripheral blood stem cell transplantation (PBSCT) has been widely used. The optimal time for collection is a critical factor to obtain proper counts of CD34 cell by peripheral blood stem cell collection (PBSC). The purpose of this study was to identify the factors influencing peripheral blood stem cell collection in order to figure out the more effective timing for PBSC. METHOD: The subjects of this study were 189 patients undergoing 3 leukapheresis from January 28, 2005 to December 31, 2006. Group's characteristics, checkup opinion of pre-peripheral blood on the day of harvest & outcome of PBSC were analyzed and evaluated using SAS statistics program after grouping patients as below; group 1-CD34 cell counts or =4x10(6)/kg (n=63). RESULTS: Based on outcome of peripheral blood stem cell according to diagnosis, acute myelocytic leukemia (AML) was 65.5% at Group 1, Lymphoma was 21.7% at Group 2 and multiple myeloma (MM) was 70.8% at Group 3. There were significant differences in CD34 cell counts according to diagnosis (p=0.00004). Type of cytokine mobilization according to diagnosis, Lenograsim was using 62.5% of MM & 38.2% of AML and filgrastim is using 22.0% of AML only. Circular peripheral blood CD34 cell counts prior to harvest was 258.1/microliter at Group 3 which was much higher comparing to Group 1 (10.5/microliter) and Group 2 (39.9/microliter) (p<0.001). TNC counts of collected peripheral blood stem cell was 15.36x10(6)/kg at Group 3 microliter and it's much higher than Group 2 (13.16x10(6)/kg) and Group 1 (12.36x10(6)/kg) (p=0.083). There was no significant difference in MNC counts inbetween 3 groups. CONCLUSIONS: Circular peripheral blood CD34+ cell counts prior to harvest was much higher at Group 3 than Group 1 and Group 2. Therefore, the number of CD34+ cells on the day of harvest can be used as an accurate predictor for peripheral blood stem cell.

The Efficacy and Safety of Large Volume Leukapheresis in Allogeneic Peripheral Blood Stem Cells Collection / 대한수혈학회지

BACKGROUND: Transplantation of allogeneic peripheral blood stem cells (PBSCs) may have advantage over bone marrow transplantation with regards to the speed of hematopoietic and immunologic recovery. Recently to overcome the need for multiple leukaphereses to collect enough PBSCs for autologous transplantation, large volume leukaphereses (LVL) are used to process multiple blood volumes per session. Experience with this technique in healthy individuals after mobilization with colony stimulating factor (CSF) is limited. We have investigated the efficacy and safety of LVL for the collection of G-CSF and GM-CSF mobilized PBSCs from healthy donors. METHODS: This study was done on 40 healthy donors who were mobilized with G-CSF and GM-CSF for allogeneic peripheral blood stem cells transplantation (allo-PBSCT). After 5 days of mobilization treatment, PBSCs were collected by LVL with Fenwal CS-3000 Plus (Baxter Co, USA). In LVL, heparin was administered in addition to ACD-A. Patients underwent of LVL procedures daily to obtain a target cell dose of >or= 4x10(8)/kg MNCs and >or= 6x10(6)/kg CD34+ cells. RESULTS: 66 LVL procedures were done on 40 donors. Of these donors, 31 (77.5%) reached the collection target with one leukapheresis. The product per LVL contained a mean 5.79+/-2.47 10(8)MNCs/kg and 11.6+/-10.62x10(6) CD34+ cells/kg respectively. Mean percentages of MNC were 79.88+/-22.15% and collection efficiencies of MNCs were inversely related to the starting MNC count (r=-0.536, P<0.001). After LVL, although none of the donors exhibited bleeding complications, platelets decreased from 187.4+/-52.68x10(3)/microL to 74.88+/-13.7x10(3)/microL and activated partial thromboplastin time (APTT) prolonged from 29.13+/-3.77 seconds to 67.51+/-54.26 seconds. CONCLUSION: We conclude that LVL after mobilization treatment with G-CSF and GM-CSF in normal healthy donors was tolerable and efficient methods for PBSCs collection, but long-term risk of adverse effects in normal donors needs to be carefully addressed by individual institutions as well as national and international registries.

Peripheral Blood Stem Cell Collection through Large Volume Leukapheresis after Mobilization with GM-CSF and/or G-CSF in Normal Healthy Donors / 대한혈액학회지

BACKGROUND: The use of colony stimulating factor (CSF) has increased to mobilize hematopoietic progenitor cells for allo-PBSCT. The most effective mobilization regimen has not yet defined. The authors analyzed the results of the mobilized PBSC collection through large volume leukapheresis from 38 normal healthy donors using three different regimens, namely, a single regimen with GM-CSF (Leucogen ), a concurrent use of GM-CSF and G-CSF (Leucostim ), and a sequential regimen with GM-CSF followed by G-CSF. METHODS: This study was done on 38 healthy donors from Sep. 1998 to Jan. 2001. One donor was mobilized with G-CSF alone, 9 with GM-CSF alone, 20 with concurrent regimen and 8 with sequential regimen. After 5 days of mobilization treatment, PBSCs were collected by large volume leukapheresis through femoral vein catheter. We compared the results of each collected progenitor cells and observed the side effects. RESULTS: The average WBC count before apheresis was 22.6+-11.0x103/uL and circulating CD34+cell percent was 1.31+-2.24%. Total 66 times with an average of 1.46+-0.61 of largevolume leukapheresis were performed for the 37 donors. The mean collected MNC count was 4.61+-2.77x108/kg, CD3+cell count was 2.95+-1.82x108/kg and CD34+cell count was 9.76+-12.42x106/kg. A significant side effect observed after large volume leukapheresis was thrombocytopenia showing decrease from 199.1+-52.2 to 80.7+-25.2x103/uL without any bleeding tendency. The mean collected MNC counts provoed to be significantly higher in combination groups with GM-and G-CSF than GM-CSF alone (P<0.05). The CD34+cell counts showed to be statistically higher in a sequential group compared to the concurrent and single regimen groups (P<0.05). CONCLUSION: A mobilization protocol with combination regimens of GM-CSF and G-CSF seemed to be superior to a single regimen with GM-CSF. Large volume leukapheresis through femoral vein catheter after mobilization with combination regimens of GM-and G-CSF in normal healthy donors was safe and proved to be an excellent. method to harvest stem cells.

Experience with 671 peripheral blood stem cell collection / 대한수혈학회지

BACKGROUNDS: Peripheral blood stem cells (PBSC) are increasingly used because of ease of collection and rapid kinetics of recovery relative to bone marrow transplantation (BMT). Here we summarize laboratory and clinical experiences of 671 PBSC harvests performed at a single institution. METHODS: PBSC harvests were performed mainly with Fenwal CS3000 plus blood cell separator. 126 cancer patients and 12 allogeneic BMT donors (54 children, 84 adults) were enrolled in this study. Total WBC and mononuclear cells (MNC) were analyzed in peripheral blood before procedure, and total WBC, MNC and CD34+ cell count on PBSC product. Correlation between dose of CD34+ cells and hematologic recovery was evaluated by Wilcoxon rank sum test and Pearson correlation analysis. RESLUTS: A mean of 4.8 leukapheresis was performed and 9,155 mL blood was processed. A mean volume of PBSC product was 66 mL and MNC purity of that was 84%. An average of 3.0 +/- 3.3x108 WBC/kg, 2.4 +/- 2.6x108 MNC/kg, and 2.7 +/- 5.4x106 CD34+ cells/kg was collected in each procedure. A goal of 1x108/kg MNCs was reached in 98.4% (127/129) of the patients and more than 1x106/kg CD34+ cells were collected in 91.8% (90/98). Various complications of PBSC harvests were observed but the incidence of serious adverse reaction was low. Median times to an absolute neutrophil count over 500/uL were 11 (7-34) and 11 (8-29) days respectively, in adult and children, and those to a sustained platelet count over 50,000/uL were 18.5 (9-118) and 36 (16-85) days, respectively. CONCLUSION: We analyzed yield, adverse reaction and hematologic recovery of PBSC harvests and this can serve basic data for the PBSC procedure.

Correlation between Progenitor Cell Dose and the Rate of Engraftment in Autologous Peripheral Blood Stem Cell Transplantation and Allogeneic Bone Marrow Transplantation / 대한수혈학회지

BACKGROUND: Bone marrow transplantation (BMT) or peripheral blood stem cell transplantation (PBSCT) following high dose chemotherapy has been an important therapeutic option for patients with hematologic malignancies or some solid tumors. The number of progenitor cells in the collection products has been used to determine the optimum time to stop the collections and to predict the hematopoietic engraftment after transplantation. In this study, we investigated the relationship between end-product cell counts measured by different methods and the influence of the infused cell dose on the engraftment rate. METHODS: Twenty five patients receiving autologous PBSCT and 25 patients receiving allogeneic BMT were studied. The number of total nucleated cells (TNC), of mononuclear cells (MNC), of CD34+ cells, and of CFU-GM (colony-forming unit-granulocyte monocyte) colonies were measured in each collection product. The number of days required to achieve an absolute neutrophil count (ANC) of 0.5x109/L with TNC count of 1.0x109/L and platelet count of 20x109/L without transfusions was taken as an arbitrary measure of the engraftment rate. RESLUTS: A close correlation between CD34+ cells/kg and CFU-GM/kg was observed in both collection products (p<0.05). However, MNC/kg also showed significant correlations with CD34+ cells/kg and CFU-GM/kg in allogeneic bone marrow collection products (p<0.05). The CFU-GM amount in the PBSC products was greater than that in the bone marrow collection products (p<0.05). Time to engraftment was a median of 14 (range 9-50) days in autologous PBSCT group, but 29 (range 17-57) days in allogeneic BMT group. In autologous PBSCT, infused CD34+ cells/kg and CFU-GM/kg correlated significantly with ANC recovery (p<0.05). CONCLUSIONS: The number of CD34+ cells was correlated with that of CFU-GM in the collection products, and the infused cell doses showed positive relation to the engraftment rate in autologous PBSCT. These findings suggest that measurement of CD34+ cell counts alone would be a sufficient parameter to predict the engraftment rate in autologous PBSCT.

Determining the Optimal Time for Peripheral Blood Stem Cell Harvest by Detecting Immature Cells using Hematology Analyzer, SE-9000TM IMI Channel / 대한임상병리학회지

BACKGROUND: A key to successful peripheral blood stem cell transplantation is to harvest a sufficient amount of hematopoietic stem cells. A method of quickly detecting hematopoietic stem cells in peripheral blood with simple procedures using the SE-9000TM IMI channel (TOA Medical Electronics Co., Ltd., Kobe, Japan) was developed. In this study, usefulness of determining the optimal time for peripheral blood stem cell harvest using IMI channel was investigated. METHODS: Seventy nine peripheral blood stem cell collections were performed from thirteen patients with hematologic malignancy and nineteen patients with solid organ malignancy. In 13 cases, G-CSF was administrered following chemotherapy. In 19 cases only G-CSF was used to mobilize the peripheral blood stem cells. The counts of leukocytes, mononuclear cells, CD34 positive cells, and IMI in peripheral blood and leukapheresis products were determined. RESULTS: The CD34 positive cell count in harvested PBSC showed positive correlation with leukocyte cell, mononuclear cell, CD34 positive cell, and IMI in peripheral blood, with correlation coefficients of 0.48, 0.27, 0.63, 0.66, respectively. Positive correlation was presented between IMI and CD34 positive cell in peripheral blood and harvested PBSC, with a correlation coefficient, 0.83 and 0.74, respectively. CONCLUSIONS: As the SE-9000TM enables determination of the number of PBSC easily and rapidly, within approximately 85 seconds, whereas CD34 assays is expensive and needs skilled operator, the measurement of IMI positive cells is clinically useful for monitoring the peripheral blood stem cell mobilization.

Preapheresis WBC, mononuclear cell and CD34+ cell counts as predictors of optimal timing of peripheral blood stem cell collection / 대한수혈학회지

BACKGROUND: Peripheral blood stem cells (PBSC) transplantation has been widely used as a substitute of bone marrow transplantation in patients with hematologic malignancies and solid tumors. Because, PBSC harvest by serial daily apheresis procedure is expensive and time consuming, it is important to determine the best time to start the collection for reducing the number of apheresis procedure. We analyzed our experiences of PBSC collections and evaluated the preapheresis hematologic parameters that may predict the PBSC yields. METHODS: One hundred seventy six PBSC harvests from seventy cancer patients (median age : 32 yrs; fourty five males and twenty five females) were performed using our large volume leukapheresis protocol (total blood volume processed : over three total blood volume) after chemotherapy and infusion of G-CSF. Peripheral blood obtained immediately before the start of apheresis was analyzed for total WBC, mononuclear cell (MNC), and CD34+ cell counts. Total WBC, MNC, and CD34+ cell count were performed on selected samples of PBSC from each patient before freezing for determining the PBSC yields. Linear regression analysis was performed on logarithmized data whether preapheresis WBC, MNC, and CD34+ cell counts on the day of harvest in the peripheral blood might correlate well with the PBSC yield, respectively. RESLUTS: With the use of linear regression analysis, preapheresis WBC counts and MNC counts were not correlated significantly with the CD34+ cell yield in PBSC harvests (WBC/microliter in PB vs. CD34+ cell/kg in harvests, r=0.35, p=0.10; MNC/microliter in PB vs. CD34+ cells/kg in harvests, r=0.42, p=0.07). But the CD34+ cell count (CD34+ cells/microliter in peripheral blood) correlated most closely with the progenitor cell yield in the corresponding leukapheresis product (CD34+ cells/kg body weight, r=0.75, p<0.001). A number of 20 circulating CD34+ cells/microliter blood ensured 2.0 x 106 CD34+ cells/kg, that is known to be a threshold dose for rapid hematologic recovery, and the best time for the collection on the same day by a single leukapheresis in more than 85% cases. CONCLUSIONS: The number of CD34+ cells/microliter blood allows a reliable prediction of the CD34+ progenitor cell yield in subsequent leukapheresis procedure, while WBC and MNC counts did not predict the progenitor cell yield. A level of more than 20 CD34+ cells/microliter indicates that the threshold quantity of 2.0 x 106 CD34+ cells/kg is likely to be obtained by a single leukapheresis processing 15~20 liters of peripheral blood.

High Dose Chemotherapy Using CD34+ Hematopoietic Stem Cells / 대한혈액학회지

BACKGROUND: Recently, CD34 antigen expressed on hematopoietic stem cells which is not detected on non-Hodgkin lymphoma (NHL), multiple myeloma and most solid tumors, is identified. In autologous bone marrow transplantation (BMT), positive selection of CD34+ cells may be used to provide hematopoietic stem cells capable of engraftment but depleted of tumor cells. And it can be used to depletion of T lymphocytes to prevent the graft versus host disease (GVHD) in allogeneic BMT. So we performed this study to evaluate the efficacy of purification of CD34+ stem cells with CEPRATE SC Stem Cell Concentration System (CellPro Inc.) and to assess the influence of CD34+ stem cells on engraftment. METHODS: Peripheral blood stem cells were mobilized with cyclophosphamide (except one patient with malignant lymphoma) and G-CSF and harvested using CS-3000 (Fenwall). CD34+ stem cells counted by FACScan (Becton-Dickinson). The conditioning regimens were ICE (Ifosphamide/Carboplatin/Etoposide) in breast cancer, high dose melphalan in multiple myeloma, BEAC (BCNU/Etoposide/Ara-C/Cyclophosphamide) in NHL, TBI (total body irradiation) with cyclophosphamide in acute lymphocytic leukemia (ALL) and busulfan with cyclophosphamide in myelodysplastic syndrome (MDS). We used G-CSF (10 microgram/kg) after transplantation in all patients. RESULTS: Eleven patients, six with high risk or metastatic breast cancer, one with refractory multiple myeloma, one with acute lymphocytic leukemia (transformed from lymphoblasticlymphoma), two with relapsed malignant lymphoma, one with myelodysplastic syndrome (HLA one-locus mismatched allogeneic BMT case, for T lymphocyte depletion) were treated. Hematopoietic stem cells were harvested from autologous peripheral blood in all patients except one patient with MDS whose stem cells were harvested from allogeneic bone marrow. Median duration and number of peripheral blood stem cell (PBSC) harvest were 15 days (13~22) and 3 times (2~8), respectively. The mean number of total stem cells and CD34+ stem cells harvested per pheresis were 204.8 (17.4~797.9)x106/kg and 3.0 (0.3~11.9)x106/kg, respectively. The mean efficacy of CD34+ hematopoietic stem cell selection by CEPRATE SC Stem Cell Concentration System was 47.7% (1.4~99.0%). The number of infused CD34+ stem cells per patient ranged from 0.34 to 4.8x106/kg (mean 2.3x106/kg). After transplantation, the median day of achieving granulocyte counts of >0.5x109/L was 10.5 days and platelet counts of >50x109/L was 14 days. CONCLUSION: CD34+ stem cells separated with CEPRATE SC Stem Cell Concentration System provided reliable and timely hematopoietic reconstitution.

Effect of hematopoietic factors mobilized peripheral blood stem cells on rhesus monkeys after irradiation of 7.0Gy ? ray / 解放军医学杂志

Objective To observe the curative effect of rhG-CSF or rhG-CSF+rhSCF mobilized peripheral blood stem cells (PBSC) on rhesus monkeys after irradiation of 7.0Gy ? ray.MethodsFifteen healthy adult rhesus monkeys were divided into placebo control, rhG-CSF, and rhG-CSF+rhSCF mobilized groups. rhSCF 20?g?kg -1 ?d -1 (?KD) was administered for 8 consecutive days. Then from the 4th day to the 8th day, animals of both rhG-CSF and rhG-CSF+rhSCF mobilized groups were also given rhG-CSF 10?KD. The placebo control animals were given 0.9% sodium chloride of the same volume per kilogram. On day 0 of the irradiation, PBSCs were collected from all subjects. 3～4 hrs after irradiation, all animals received autologous PBSC infusion. ResultsFrom day 21 to day 29 after TBI, rhG-CSF+rhSCF mobilized group showed higher WBC counts compared to placebo control and rhG-CSF mobilized group. Platelet counts of both mobilized groups recovered more quickly than those of placebo control. Bone marrow nucleated cells culture demonstrated that rhG-CSF and rhG-CSF+rhSCF mobilized PBSC had stimulated bone marrow nucleated cells to form more CFUs. Histopathological evaluation showed the number of hematopoietic cells in the bone marrow of rhG-CSF mobilized group was greater than those in placebo control but less than those in rhG-CSF+rhSCF mobilized group. ConclusionThe infusion of rhG-CSF or rhG-CSF+rhSCF mobilized PBSC can improve the hematopoietic recovery in rhesus monkeys after 7.0Gy ? ray irradiation. The combined use of two mobilizing factors gives better result than using single factor.

THE AFFECT OF THE ABO BLOOD GROUP ON THE HEMATOPOIETIC RECONSITITUTION IN ALLO-PBSCT FOR MALIGNANCE HEMATOLOGICAL DISEASE / 解放军医学杂志

0 05). The PBSC with red blood cells(8～11ml) without elimination before infusion were harvested by CS 3000 Plus.The leve1 of blood group antigens and antibodies and bone marrow cells morphology were examined at different intervals after transplantation. No patient showed acute haemolysis in the ABO incompatible group. The median time of blood type transformation was 40 2?25 days. The median time for recovery of normal WBC and Plt in ABO incompatible groups was 10 days longer than that in the ABO compatible group. The number of patients with Hb