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【Objective】 To evaluate the safety and efficacy of the collection of peripheral blood stem cells (PBSCs) in pediatric patients with thalassemia major (TM) weighing 20 kg or less. 【Methods】 PBSCs collection data of 170 pediatric patients with TM weighing 20 kg or less from January 2013 to December 2020 in our center were reviewed. Safety was assessed by the occurrence of adverse events during apheresis procedures, and efficacy was evaluated by the number of CD34+ cells collected. 【Results】 A total of 171 PBSCs procedures were performed on 170 patients with TM weighing 20 kg or less, with a median age of (4.98±1.53) years and a median weight of (17.30±2.18) kg. The probability of collecting at least 1×106 CD34+ cells/kg during a single course of apheresis was 99.41% (169/170), with a median (5.88±4.23) ×106 CD34+ cells collected per kg of weight of the recipient. A minimum pre-apheresis hemoglobin (Hb) of 60 g/L in patients with TM weighing 20 kg or less was safe and feasible. The most common adverse event of G-CSF mobilization in TM patients is bone pain, with the incidence of 7.65% (13/170), which was higher than that of healthy children donors in our center. The most common adverse events during the collection were pain at the puncture site of the femoral vein (6.47%, 11/170) and low pressure of the fluid (2.92%, 5/170). And no serious complications related to PBSCs mobilization, central venous catheter(CVC)placement or the apheresis procedure occurred. 【Conclusion】 PBSCs collection by COM.TEC blood cell separator in children weighing 20 kg or less is safe and efficacious.
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Abstract Introduction The isolation of captured peripheral blood mononuclear cells (PBMNCs) from leukoreduction filters (LRFs) can be of great importance in terms of bringing the lost cells back into use. Objective The aim of this study was to evaluate various methods based on their potential to recover the peripheral blood cells from LRFs with a focus on mononuclear cells (MNCs). Method For cell isolation from LRFs, three distinct methods (back-flushing, direct and vacuum pump) were compared through the calculation of the yield of isolated MNCs. The viability of extracted cells was determined by the flow cytometry technique. Moreover, the recovered MNCs were characterized regarding the presence of blood stem cell purification. The cell culture, microscopic observation, and immunophenotyping were employed to characterize the blood stem cells (hematopoietic, mesenchymal and progenitor endothelial stem cells). Results The yield of isolation obtained in the back-flushing, direct and vacuum pump methods were 17.7 ± 1.28, 17.3 ± 0.96 and 21.2 ± 0.90 percent, respectively. Although the highest potential for total blood cell recovery belonged to the vacuum pump method, the lowest cell viability (85.73 ± 4.84%) was observed in this method. However, the isolation process of the back-flushing and direct methods had less effect on cell viability. The characterization of the isolated MNCs displayed that the dominant positive phenotype was for CD34/CD45, indicating hematopoietic stem cells. In addition, the endothelial stem/progenitor cells were significantly detected as CD31/CD133 positive cells. Conclusion According to our results and considering the safety and efficiency potential of each of the applied methods, the back-flushing in comparison with the other methods can be considered a suitable procedure for MNC isolation from LRFs.
Subject(s)
Leukocytes, Mononuclear , Cell Separation , Peripheral Blood Stem Cells , Blood Cell Count , Flow CytometryABSTRACT
@#The treatment of chronic Achilles tendinopathy (CAT) remains challenging. We report three cases of CAT treated with autologous peripheral blood stem cells (PBSCs), following principles developed for chondrogenesis of the knee joint. Outcome measurement with a minimum of one and a half years follow-up showed significant improvement of Victorian Institute of Sport Assessment-Achilles questionnaire (VISA-A) scores, with reduction of tendon thickness and inflammation on MRI scan.
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@#Osteochondral lesions of the talus (OLTs) may progress to ankle arthritis needing ankle arthroplasty or arthrodesis. We report five cases of OLTs treated along the principles developed for chondrogenesis of the knee joint with autologous peripheral blood stem cells (PBSCs), resulting in repair and regeneration of the bone and cartilage components. Improvement in Ankle Osteoarthritis Scale (AOS) scores with minimum two years follow-up showed statistical significance (p < 0.05).
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@#End-stage ankle arthritis represents an “unmet medical need”, awaiting an appropriate time for joint arthroplasty or arthrodesis. We report three cases of end-stage ankle arthritis treated along the principles developed for chondrogenesis of the knee joint with autologous peripheral blood stem cells, resulting in reversal of the ankle arthritis. The improvement in clinical outcome measure scores (Ankle Osteoarthritis Scale total score) with a minimum two-year follow-up were comparable to total ankle replacement (TAR), arthroscopic ankle arthrodesis (AAA) and open ankle arthrodesis (OAA).
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@#This is a case report of a Gustilo-Anderson Type IIIB comminuted open right tibial fracture with massive bone loss, complicated by methicillin-resistant Staphylococus aureus (MRSA) infection. Non-viable and contaminated bony fragments were removed and infected bone resected. Soft tissue coverage and antibiotics were effective against the MRSA infection. A unifocal bone transport with the Ilizarov method regenerated 13cm of the missing tibia. Autologous peripheral blood stem cells (PBSC) injections into the osteogenesis site boosted bone regeneration and consolidation with a shortened Bone Healing index (BHI) of 23 days/cm.
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The efficacy and safety of co-transplantation of unrelated donor peripheral blood stem cells (UD-PBSCs) combined with umbilical cord mesenchymal stem cells (UC-MSCs) in refractory severe aplastic anemia-Ⅱ(RSAA-Ⅱ) were analyzed retrospectively. Fifteen patients with RSAA-Ⅱ underwent UD-PBSCs and UC-MSCs co-transplantation, among whom 14 cases had hematopoietic reconstitution without severe graft versus-host disease (GVHD). The 5-year overall survival rate was 78.57%. Combination of UD-PBSCs and UC-MSCs transplantation could be a safe and effective option for RSAA-Ⅱ.
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ObjectiveTo investigate the efficiency of autologous transplantation of peripheral blood stem cells for treatment of patients with diabetic lower limb ischemia.MethodsEighteen patients of type 2 diabetes with diabetic lower limb ischemia (30 legs) were treated by autologous transplantation of peripheral blood stem cells.ResultsThe limb pain, cool feeling and numbness feeling improved significantly after PBSC transplantation,the improvement rate were 96.7％, 100.0％ and 95.8％ respectively.Intermittent claudication was also relieved significantly, total remission rate was 76.9％.The ABI and TcPO2 of patients increased significantly at 3 months after transplantation.After the transplantation ABI raised from 0.60 ± 0.11 to 0.71 ±0.12(t =-6.882, P ＜ 0.01) .93.3％ of patients' TcPO2 raised in different degrees.The foot infections were well controlled.Ulcer or toes gangrene got better or healed.No obvious complications or adverse reaction were observed after the transplantation.ConclusionAutologous transplantation of peripheral blood stem cells shows to be a simple, safe and effective method in treating patients with diabetic lower limb ischemia.
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Human tumors, including those of the hepatobiliary system, express a number of specific antigens that can be recognized by T cells, and may provide potential targets for cancer immunotherapy. Dendritic cells (DCs) are rare leucocytes that are uniquely potent in their ability to capture, process and present antigens to T cells. The ability to culture sufficient numbers of DCs from human bone marrow or blood progenitors has attracted a great deal of interest in their potential utilization in human tumor vaccination. CD34+ peripheral blood stem cells (PBSCs) were obtained from a patient with a hepatocellular carcinoma. The PBSCs were cultured in the X-VIVO 20 medium supplemented with the Flt-3 Ligand (FL), GM-CSF, IL-4 and TNF-alpha for 12 days. The morphology and functions of the cells were examined. The generated cells had the typical morphology of DCs. When the DCs were reinjected into the same patient, an augmentation of the cytotoxic T lymphocyte (CTL) activity was observed. Concomitantly, an increase in the natural killer (NK) cell activity was also detected in the patient. These results suggest that DCs-based cancer immunotherapy may become an important treatment option for cancer patients in the future.
Subject(s)
Humans , Bone Marrow , Carcinoma, Hepatocellular , Dendritic Cells , Granulocyte-Macrophage Colony-Stimulating Factor , Immunotherapy , Interleukin-4 , Lymphocytes , Membrane Proteins , Monitoring, Immunologic , Stem Cells , T-Lymphocytes , Tumor Necrosis Factor-alpha , VaccinationABSTRACT
BACKGROUND/AIMS: Peripheral blood progenitor cells (PBSC) have been used instead of bone marrow cells for allogeneic transplantation. Progenitor cells are mobilized using granulocyte colony stimulating factor (G-CSF). Kinetic studies of G-CSF have shown that the CD34+ cells peak on day 5, although the optimal time for harvesting has not been established. METHODS: We compared two mobilization schedules using G-CSF. In groups I and II, G-CSF 10 microgram/kg/day was administered subcutaneously for 3 or 4 days, respectively, and leukapheresis was performed the next day. RESULTS: In groups I and II, a respective mean number of 5.84 x 106 and 10.76 x 106 circulating CD34+ cells per kilogram of the recipient's weight was collected by leukapheresis. The yield of the two collections did not differ significantly (p=0.386). CONCLUSION: This study suggested that the two mobilization schedules are equally effective. To avoid prolonged exposure to G-CSF, the day 4 collection schedule is the preferred mobilization strategy.
Subject(s)
Humans , Appointments and Schedules , Bone Marrow Cells , Colony-Stimulating Factors , Granulocyte Colony-Stimulating Factor , Granulocytes , Leukapheresis , Stem Cells , Tissue Donors , Transplantation, HomologousABSTRACT
Objective To analyse the effect of mobilization and collection's time of peripheral blood stem cells(PBSC) from 8 cases of healthy donors. Methods The 10 donors were studied by self-control design.The number of aphereses was two times every donors. Healthy donors received rhG-CSF according to two different PBSC collection starting time: group 1:PBSC collection was starts 2 hours(2 h) after the fourth day or the fifth day of rhG-CSF. group 2:PBSC collection was starts 4 hours(4 h) after the fourth day or the fifth day of rhG-CSF.(The first dose of rhG-CSF was given on day 1, considering day 0 as the day before starting mobilization). In this study we have compared with two groups of apheresis product. Results The MNC count was significantly higher for donors 4 h collection (groups 2) then 2 h. ( groups 1)(P
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Allogeneic peripheral blood stem cells transplantation is the most effective method for maligant hematological disease. The author reviewed the indication of acceptance of HLA mismatch kinds and the prediction of the results by selecting the measures such as pretreatment,mobilization,stem cells disposal,immunity reconstruction,graft failure management in order to improve the engraft rate.
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BACKGROUND: An essential prerequisite for successful procurement of sufficient peripheral blood stem cells (PBSC) for engraftment is the optimal timing of collection. The Sysmex SE-9000 automated hematology analyzer provides the immature information (IMI) channel for the identification and counting PBSC. In this study, The optimal timing of PBSC collection was studied using IMI channel. METHODS: 193 peripheral blood stem cell collections were performed from 52 patients with hematologic disorders or solid tumors and 15 donors. Pre-harvest peripheral blood WBC, mononuclear cells (MNC) and IMI were tested and compared with CD34+ cell count and CFU-GM count of harvested products. RESULTS: Peripheral blood WBC and MNC count showed a weak correlation with CD34+ cell yield (r=0.38, P1x10(6)/kg with sensitivity of 88.7%. Positive and negative predictive values of IMI >465/microliter for CD34+ cell >1x10(6)/kg were 65.5% and 87.5%, respectively. CONCLUSIONS: The automated IMI might be used as a simple and efficient indicator of PBSC mobilization and applying variable cutoff values of IMI would be a useful tool to predict the optimal timing of PBSC collection.
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Humans , Cell Count , Granulocyte-Macrophage Progenitor Cells , Hematology , ROC Curve , Stem Cells , Tissue DonorsABSTRACT
Objective To find a safe and effective way for the collection of stem cells from peripheral blood in low body weight children. Methods Peripheral blood stem cells were collected altogether 15 times in 5 children with body weight lower than 20 kg. One day before the collection, a double cavity tube specially made for children was inserted into femoral vein and remained there for future use. Peripheral blood stem cells were separated by the separation vessel specially designed for children and collected in the collecting tank for children. After initialization, alien blood irradiated by X ray was infused in and moved through the separation machine to keep the blood balanced and hematocrit stable in the body of children. During the separation, anticoagulant/whole blood was kept between 1∶11 and 1∶13. The volume of blood processed every time was 2-4 times of total volume of child blood(2 700-5 850 ml). Results Mononuclear cells and CD34 + cells were successfully collected each time in all 5 children, enough for the transplantation of peripheral blood stem cells. Conclusion The way we developed to collect peripheral blood stem cells in children with low body weight is safe and effective.
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We investigated the effect and outcome of allogeneic peripheral blood stem cell (PBSC) rescue for aplastic anemia (AA) patients with graft failure after allogeneic bone marrow transplantation (BMT). Seven (28%) of 25 AA patients who received BMT from HLA-identical sibling donors developed late graft failure at a median of 7 months (range, 2.0-9.3 months) after transplantation. The patients with graft failure were treated with PBSC collected from the original donor after mobilization with granulocyte-colony stimulating factor (G-CSF). The median boost dose of peripheral blood mononuclear cells was 3.1 x 10(8)/kg (range, 1.4-11.9 x 10(8)/kg). Median times to reach an absolute neutrophil count greater than 0.5 x 10(9)/L and a platelet count greater than 50 x 10(9)/L were 7 days (range, 4-14 days) and 9 days (range, 3-41 days), respectively. There was sustained graft function in 6 of 7 patients, with a median follow-up duration of 3.3 yr (range, 1.0-6.2 yr). Grade-I acute graft-versus-host disease (GVHD) occurred in 2 patients, while extensive chronic GVHD developed in 3 patients. This report shows that G-CSF-mobilized allogeneic PBSC rescue is very effective in achieving complete and sustained engraftment in patients with AA after graft failure. However, more efficacious measures to prevent extensive chronic GVHD remain to be developed.
Subject(s)
Adolescent , Adult , Child , Child, Preschool , Female , Humans , Infant , Male , Anemia, Aplastic/therapy , Bone Marrow Transplantation , Graft Survival , Graft vs Host Disease , Granulocyte Colony-Stimulating Factor/administration & dosage , Hematopoietic Stem Cell Transplantation , Hematopoietic Stem Cells/drug effects , Survival Rate , Transplantation, Homologous , Treatment OutcomeABSTRACT
To evaluate the effects of recombinant human stem cell factor on mobilization of peripheral blood stem cells, 30 monkeys were divided into control, rhG CSF 10?g?kg -1 ?d -1 (?KD), rhSCF 50?KD, rhG CSF 10?KD+rhSCF 20?KD, rhG CSF 10?KD+rhSCF 50?KD and rhG CSF 10?KD+rhSCF 125?KD groups. The monkeys were given subcutaneous injections of rhSCF for 8 days and/or rhG CSF for 5 days. The highest value of leukocyte in rhG CSF and rhSCF groups was 206% and 200% respectively, and that of rhG CSF+rhSCF groups was 280% to 310%. CFU GM of peripheral blood was up to 1 1, 1 3 and 5 8～7 9 times respectively after the mobilization. CD34 positive cell number of joint mobilization group was 20, 24 and 35 times respectively of that before mobilization. rhSCF can mobilize the effect of peripheral blood stem cells, and the effect of joint rhSCF+rhG CSF is better.
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Objective To evaluate the effects of recombinant human hematopoietic synergistic factor (rhHSF) on rapid mobilization of the peripheral blood stem cells. Methods 8 monkeys were divided into rhG-CSF 10?g/(kg?day) and rhG-CSF 10?g/(kg?day) + rhHSF 500?g/(kg?day) group, with 4 monkeys in each group. All of them received rhG-CSF subcutaneously once daily for 4 days, and in the latter group, the animals received rhHSF subcutaneously in single dose on the fifth day. Results The highest counts of CD34~+ cell, CFU-GM and neutrophils in animals receiving rhHSF only were 23, 1.8 and 3.3 times of baseline value at 180, 45 and 60 min, while those in animals receiving rhG-CSF and rhHSF were 41.3, 8.3 and 4.9 times of base line value. Conclusion rhHSF can rapidly mobilize hematopoietic stem cells and neutrophils into peripheral blood, and it is found to synergize with G-CSF to augment stem cell mobilization, suggesting a potent clinical utility of rhHSF in peripheral blood stem cells transplantation.
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Purpose:To evaluate the effect of Auto-PBSCT combined with long-time low-dose interleukin-2 (after transplant) for increasing the rate of 5-year disease-free survival of lymphoid malignancies (ALL and NHL).Methods:U- sing chemotherapy combined with low dose G-CSF to mobilize peripheral blood stem cells,we carried out auto-PBSCT in lymphoid malignancies (7 cases CR_1 ALL、1 case CR_2 ALL、3 cases CR_1 NHL、5 cases refractory NHL).After transplant, 15 cases used low dose interleukin-2 at 5.10~5U/d for one year or longer.Results:7 cases CR_1 ALL are still alive,the aver- age disease-free survival is 73 months.Median disease-free survival is 51.5 months.2 of 3 cases CR_1 NHL are still alive. The 6 years disease-free survival in 10 cases CR_1 lymphoid malignancies(CR_1 NHL and CR_1 ALL) is 0.90?0.11,the aver- age disease-free survival is 66.8 months.In another PR (refractory) group of NHL and ALL (CR_2),3 years disease-free survival was 0.33?0.18,and average survival time was 20.66 months,but a chemotherapy-sensitive NHL is still alive (63 months),a case of lymphoblast cell NHL also survived for 46 months and died after relapse.Conclusions:Auto-PBSCT combined with long-turm low dose IL-2(after transplant) could effectively increase the rate of 5-year disease-free survival in CR_1 lymphoid malignancies(ALL and NHL).
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BACKGROUND: There are highly-polymorphic DNA markers in the human genomic DNA, known as the variable number of tandem repeats (VNTR). The VNTR markers can be used to evaluate the engraftment of stem cells. We evaluated the discrimination power of 3 types of long-tandem repeat (LTR) and tried to predict underlying disease relapses by DNA chimerism. METHODS: Twenty-nine patients were transplanted with allogeneic peripheral blood stem cells (PBSCs) and their related donors were tested. We used the three long-tandem repeats (LTR) D1S80, D1S111, and YNZ22 for VNTR-PCR. The informative test was performed before transplantation. The chimerism analysis was performed on days +30, +60, +90, and +180 after transplantation. RESULTS: The most informative marker was D1S80 with 55.2% discrimination potential. The power of discrimination was 79.3% in the combination of 3 LTRs. Twenty-two cases were tested for DNA chimerism analysis. When the complete chimerism was represented, the engraftment was more successful and when the mixed chimerism was represented, the underlying disease relapse rate increased. CONCLUSTIONS: DNA chimerism analysis was useful to evaluate the marrow status of patients. It also served as an indication for donor lymphocyte infusion. However, compared to unrelated allogenic PBSCT, the discrimination potential for the combination of 3 LTR loci was lower in the related allogenic PBSCT. Therefore, it is thought that additional short-tandem repeats and DNA sequencing are required for more discrimination power especially in related transplantation cases.
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Humans , Bone Marrow , Chimerism , Discrimination, Psychological , DNA , Genetic Markers , Lymphocytes , Minisatellite Repeats , Recurrence , Sequence Analysis, DNA , Stem Cells , Tissue DonorsABSTRACT
BACKGROUND: The optimal timing of peripheral blood stem cell (PBSC) collection is essential to successful procurement of sufficient PBSC for engraftment. The purpose of this study was to evaluate the peripheral blood parameters that may predict the apheretic yield of circulating stem cells in patients with breast cancer. METHODS: We did a retrospective review of 29 patients with breast cancer (14 : high risk, 15 : metastatic disease) who underwent mobilizing therapy from Dec. 1992 to Jan. 1999. Immediately prior to 119 consecutive PBSC collection procedures, the PB white blood cell (WBC) and monocyte were determined and correlated with stem cell parameters namely, CD34+ cell and mononuclear cell content. RESULTS: The median of 0.57x106CD34+cells/kg patient body weight (range, 0-9.39) were collected per harvest. The WBC on the day of apheresis showed only weak correlation with the mononuclear cells collected (r=0.26). In contrast, the WBC count and monocyte count in PB did not correlated with CD34+ cells harvested CONCLUSION: WBC and monocyte count are not appropriate parameters to identify the exact timing for apheresis and predict the amount of peripheral blood stem cells collected in patients with breast cancer.