Purpose: Long-term culture- initiating cells(LTC-IC) are stem cells that have the capacities of long-term engraftment and helping to establish hematopoietic microenvironment. For evaluation of the LTC-IC, we measured the counts and function with multidimentional flowcytometry in long-term culture media. METHODS: Samples were obtained from umbilical cord blood, leukapheresis products and bone marrow(BM). LTC-IC were counted with flowcytometric analysis using anti- CD34, anti-CD38, and anti-HLA-DR antibodies at 0, 5, and 8 weeks. Cell adhesion molecule related with stem cell were evaluated with flowcytometric analysis also using anti-VCAM-1(CD106) and anti-VLA-4(CD49d) at 0 and 8 weeks. RESULTS: The proportion of CD34+/CD38- cell from fractionated mononuclear cells at 0 week were 0.46%, 0.044%, and 0.038% for BM, leukapheresis products, and umbilical cord blood respectively and then rapidly decreased at 5 weeks, but still persisted at 8 weeks in all three groups. The proportion of CD34+/HLA-DR- cells was the same tendency to CD34+/CD38-. VCAM+ expression rate from fractionated CD34+ cells at 0 and 8 weeks were 67.3% and 40.2% for BM and 64.1% 44.2% for umbilical cord blood but it was very low 31.2% and 5.1% for leukapheresis products. VLA-4+ expression rate for fractionated CD34+ cells at 0 and 8 weeks were similar tendency to VCAM+ cells. CONCLUSION: This study suggest that the count of LTC-IC decreased with time but still persisted until 8 weeks. Umbilical cord blood including BM help to establish the hematopoietic microenvironments.
Antibodies ; Cell Adhesion ; Culture Media ; Fetal Blood ; Leukapheresis ; Stem Cells

This study was aimed to investigate the effect of leukocyte depletion by filtration on the quality of apheresis platelets. 20 units of donor apheresis platelets were randomly selected and were preserved with agitation at 20 - 24 degrees C for 24 - 96 hours, then were filtered on polyester flatbed filters. The platelet concentration, mean platelet volume (MPV), volume of apheresis platelets, leukocyte count, pH value, lactate dehydrogenase (LDH) concentration, K(+) concentration and CD62p expression level on surface of platelet membrane, were detected before and after filtration, as well as the rate of leukocyte depletion and platelet loss were calculated. The results showed that the leukocyte count after filtration was remarkably lower than that before filtration (p < 0.001), and the rate of leukocyte depletion was 99.97%. Platelet loss was approximately 8%, and obviously lower than that of the national standard (p < 0.001). MPV, pH value, K(+) and LDH concentration were not significantly different before and after filtration. Compared with platelets before filtration, CD62p expression level after filtration slightly decreased (p > 0.05). CD62p expression on surface of platelet membrane in perfusion fluid obtained from filter plate was obviously higher than that before filtration (p < 0.05). MA of platelet after filtration slightly decreased (p > 0.05). It is concluded that leukocyte and partial activated platelets can be removed efficiently by using polyester flatbed filters, and platelet loss is very low. Filtration does not adversely affect coagulation activity of the platelets in vitro. Apheresis platelets after filtration can fulfil quality requirements to prevent infection of cytomegalovirus and HLA alloimmunization.
Filtration ; Humans ; Leukapheresis ; instrumentation ; Platelet Count ; Plateletpheresis ; instrumentation ; methods

OBJECTIVE: To improve the collection efficiency of leukapheresis, explore relatively scientific and objective evaluation indicators for collection effect, and observe the effect of high-volume leukapheresis on blood cells and coagulation function. METHODS: A total of 158 times of high-volume leukapheresis were performed on 93 patients with hyperleukocytic leukemia by using continuous flow centrifugal blood component separator. 1/5-1/4 of total blood volume of the patients was taken as the target value of leukocyte suspension for single treatment. In addition, the total number of white blood cells (WBCs) subtracted, value of WBCs reduction, rate of WBCs reduction, decrease value of WBCs count, decrease rate of WBCs count, amount of hemoglobin (Hb) lost, value of Hb lost, decreased value of Hb, total number of platelet (PLT) lost, the value of PLT loss, and decrease value of PLT count were used to comprehensively evaluate the collection effect of leukapheresis and influence on Hb level and PLT count of the patients. The prothrombin time (PT), activated partial thromboplastin time (aPTT), thrombin time (TT), and fibrinogen (Fib) concentration were detected before and after treatment, and the effect of leukapheresis on coagulation function of the patients was observed. RESULTS: The volume of leukocyte suspension collected in a single treatment was 793.01±214.23 ml, the total number of WBCs subtracted was 353.25 (241.99-547.28)×10⁹, the value of WBCs reduction was 86.98 (63.05-143.43)×10⁹/L, the rate of WBCs reduction was 44.24 (28.37-70.48)%, decrease value of WBCs count was 65.73 (37.17-103.97)×10⁹/L, decrease rate of WBCs count was (35.67±23.08)%, the amount of Hb lost was 17.36 (12.12-24.94) g, the value of Hb lost was 4.31 (3.01-6.12) g/L, decreased value of Hb was 4.80 (-1.25-9.33) g/L, total number of PLT lost was 222.79 (67.03-578.31)×10⁹, the value of PLT loss was 54.45 (17.29-139.08)×10⁹/L, and decrease value of PLT count was 26.00 (8.38-62.50)×10⁹/L. Before and after a single treatment, the PT was 14.80 (13.20-16.98) s and 15.20 (13.08-16.90) s (z=-1.520, P>0.05), the aPTT was 35.20 (28.68-39.75) s and 35.40 (28.00-39.75) s (z=-2.058, P<0.05), the TT was 17.50 (16.30-18.80) s and 17.70 (16.70-19.10) s (z=-3.928, P<0.001), and the Fib concentration was 2.87±1.13 g/L and 2.64±1.03 g/L (t=7.151, P<0.001), respectively. CONCLUSION High-volume leukapheresis can improve the efficiency of leukapheresis while maintaining the relative stability of the patients' circulating blood volume. The degree of influence on the patients' Hb level, PLT count, Fib concentration, and comprehensive coagulation indicators reflecting the patients' intrinsic and cxtrinsic coagulation activity is within the body's compensation range.
Blood Coagulation Tests ; Fibrinogen ; Hemoglobins ; Humans ; Leukapheresis ; Leukemia

Therapeutic leukapheresis is a treatment modality which selectively remove abnormal leukocytes and relieve symptoms produced by abnormally high concentration of leukocytes in the blood stream. Authors report the experiences of therapeutic leukapheresis at Seoul National University Hospital during the past 7 years(1988 to 1995). A total of 48 procedures were done for 29 patients(24 males and 5 females). The age distribution was from 16 years to 60 years and mean age was 30 years. The most common diagnosis of patients were acute lymphoblastic leukemia(7/15, 46.7%) and major indications for therapeutic leukapheresis were dyspnea, headache, abdominal pain and mental alteration. The mean leukocyte count before and after leukapheresis were 277,300/micro liter and 220,700/micro liter, respectively(20% decrease, P<0.001). Hemoglobin concentration was also decreased from 7.7g/dL to 7.1g/dL after leukapheresis(P<0.01). The mean number of leukocytes removed per procedure was 3.1x1011. Adverse reactions such as abdominal pain and dizziness were observed in two cases. The leukocyte count was decreased in all patients and improvement of symptoms was observed in four patients. In conclusion, therapeutic leukapheresis is relatively safe and can be used to relieve leukostatic symptoms in leukemic patients.
Abdominal Pain ; Age Distribution ; Diagnosis ; Dizziness ; Dyspnea ; Headache ; Humans ; Leukapheresis* ; Leukocyte Count ; Leukocytes ; Male ; Rivers ; Seoul

BACKGROUND: Dendritic cells (DCs) are the most potent stimulators of immune response including antitumor response. DCs are currently being pursued clinically in the development of cancer vaccines; therefore there are demands for large-scale and clinical-grade generation of DCs. In the present study, to find out the most efficient separation method of DC precursors, we compared two separation methods, namely, based on magnetic based selection and plastic adherence selection. METHODS: MNCs were collected by leukapheresis from healthful donors and separated by CD14 + immunomagnetic adsorption or plastic adherence. DC precursors separated using the two methods were differenciated in the same condition. Matured DCs were compared in terms of yield, viability, the expression of surface markers and ability to induce immune reaction. RESULTS: This study demonstrated that mature DCs from CD14 + monocytes separated using CD14 + immunomagnetic adsorption had higher expression of surface markers of DCs, yield (1.9 +/-0.5% vs. 0.5 +/-0.2%), viability (94.7 +/-2.5% vs. 72.8 +/-7.5%) and better functionality in inducing immune reaction than those from plastic adherent cells. CONCLUSION: These results demonstrated that CD14 + immunomagnetic adsorption was found to be more effective than the adherent selection for the generation of DCs. This study will allow researcher to facilitate choosing the appropriate protocol to obtain DCs.
Adsorption* ; Cancer Vaccines ; Dendritic Cells* ; Humans ; Leukapheresis ; Monocytes* ; Plastics ; Tissue Donors

BACKGROUND: Recently, the commercial kits for measurement of the absolute number of CD34+ cells have been introduced as a standard method. The aims of this study was to investigated optimal timing of peripheral blood progenitor cell(PBPC) collection and optimal CD34+ cells dose transplanted by measurement of the absolute CD34+ cells. METHODS: We measured total leukocyte count, mononuclear cell count and the absolute number of CD34+ cells using ProCOUNT(Becton Dickinson, USA) in peripheral blood from 54 patients and 7 normal donors who underwent 101 leukapheresis for PBPC collection. We studied correlations among the absolute number of circulating CD34+ cells, other predictors and harvesting yields. We investigated relationships between the posttransplant hematopoietic recovery and CD34+ cells dose in 30 patients. RESULTS: The total number of CD34+ cells in harvesting products could be mostly predicted from the absolute number of circulating CD34+ cells. From 4 to 6 day after G-CSF mobilization, the absolute number of circulating CD34+ cells was peaked. A number of circulating CD34+ cells more than 20/microliter ensured 2.5x106 CD34+ cells/Kg in harvesting products. The patients received CD34+ cells dose >3.5x106/Kg led to a significantly faster recovery of platelets, compared with the patients receiving <3.5x106 CD34+ cells/Kg(P<0.05). CONCLUSIONS: These results suggest that PBPC collection should be started at day of circulating CD34+ cells more than 20/microliter or 4-6 days after G-CSF mobilization for successful leukapheresis and the CD34+ cell dose more than 3.5x106/Kg for PBPC transplantation could predicted rapid hematopoietic recovery.
Cell Count ; Granulocyte Colony-Stimulating Factor ; Humans ; Leukapheresis ; Leukocyte Count ; Stem Cells* ; Tissue Donors

PURPOSE: Cryopreservation has been the standard method of storing hematopoietic cells for the past 20 years, but this prdegrees Cedure is laborious and expensive. So, we evaluated the hematopoietic recovery of stored PBSCs at 4degrees C for a variable storage period MATERIALS AND METHODS: Eight leukapheresis products were kept unprdegrees Cessed at 4degrees C for 96 hours. To evaluate the effect of storage period on the hematopoietic recovery of PBSCs, assays for viability of mononuclear cells (MNCs), CFU-GM colony counts and CD34 cell counts were performed every 24 hours after PBSC collection. We tried to compare hematopoetic recovery of stored PBSCs at 4degrees C with that of cryopreserved PBSCs by using repeated measures ANOVA. RESULTS: Viability of MNCs, CFU-GM colony counts and CD34 cell counts were monitored at 24 hour, 48 hour, 72 hour and 96 hour after PBSC collection. Data are expressed as percentage of baseline value and shown as mean s.d.; MNCs viability (96+/-2%, 94+/-2%, 92+/-2%, 88+/- 3%), CFU-GM colony counts (87+/-10%, 79+/-11%, 65+/-13%, 56+/-15%), and CD34 cell counts (93+/-13%, 93+/-12%, 88+/-14%, 85+/-19%). After storing PBSCs at 4degrees C for 96 hours, viability of MNCs and CFU-GM colony counts were significantly reduced (p<0.05) except CD34 cell concentration (p>0.05). Prdegrees Cedures of controlled-rate freezing and thawing resulted in a notable loss of viability (77+/-9%) and CFU-GM colony count (71+/-29%). CFU-GM colony counts of 72 hour-stored PBSCs at 4degrees C was similar to those of cryopreserved PBSCs. CONCLUSION: If G-CSF mobilized PBSCs are stored at 4degrees C in less than 72 hours after collection, those hematopoietic recovery would be comparable to that of cryopreserved stem cells which are achieved by the rate-control freezer.
Bezafibrate ; Cell Count ; Cryopreservation ; Freezing ; Granulocyte Colony-Stimulating Factor ; Granulocyte-Macrophage Progenitor Cells ; Leukapheresis ; Stem Cells*

BACKGROUND/AIMS: In chronic inflammatory conditions such as ulcerative colitis (UC), the migration of granulocytes and monocytes/macrophages from the circulation into the colonic mucosa is especially important in maintaining inflammation. The aim of this trial was to assess safety and efficacy of granulocyte and monocyte adsorption apheresis in patients with moderate-to-severe UC refractory to conventional drug therapies. METHODS: Twenty-seven patients with moderate (55.6%) to severe (44.4%) active UC refractory to conventional drug therapies who had no changes in their conventional therapy regimen in the past two weeks before the recruitment were enrolled in an open-label trial. Concomitant medications were allowed, and steroids were tapered down according to the clinical activity during the course. We used an adsorptive type extracorporeal column (Adacolumn(R); JIMRO, Takasaki, Japan), which selectively adsorb granulocytes and monocytes. Patients took five apheresis sessions, each with 60 minutes duration for 5 consecutive weeks. The primary efficacy variables were clinical disease activity, short inflammatory bowel disease questionnaire (SIBDQ), C-reactive protein (CRP), and endoscopic scores. These variables were scored at regular intervals, and analyzed at week 7 on an intention-to-treat (ITT) principles. RESULTS: At 7 weeks, 70.4% of patients showed overall improvement. Clinical disease activity (p<0.0001), endoscopic score (p<0.001), and the quality of life as assessed by SIBDQ (p<0.0001) were significantly improved after the therapy. In 56.3% of concomitant steroid users, tapering down or discontinuation of steroids was possible. Treatment was well tolerated, and no severe adverse events were observed. CONCLUSIONS: Adacolumn was very efficacious in patients with moderate-to-severe active UC refractory to conventional drug therapy, but further assessment is needed.
Adult ; Colitis, Ulcerative/*therapy ; English Abstract ; Female ; *Granulocytes ; Humans ; *Leukapheresis ; Male ; *Monocytes

BACKGROUND/AIMS: Cytapheresis (CAP) is a novel strategy for ulcerative colitis (UC). However, there is insufficient data on the long-term outcome of UC patients who achieve remission by CAP. This study involved patients with severe UC who refracted to intravenous (iv) corticosteroid. METHODS: Forty-seven UC patients who had received CAP therapy for the first time within 1 year after UC diagnosis were followed for 36 months. One of the inclusion criteria was a clinical activity index (CAI) of > or =7 points at the end of a 2-week iv course of corticosteroid therapy. CAP therapy consisted of ten sessions over 10 weeks. RESULTS: CAP induced clinical remission (CAI< or =4) in 70.2% patients (33/47). The number of submissions for colectomy was higher for severe UC at entry (CAI> or =12, n=25) than for moderately severe UC at entry (7< or =CAI<12, p=15; p<0.02). The cumulative rates of avoiding surgery and relapse were 54.5% and 24.2%, respectively, at 36 months in patients who responded to CAP therapy. This was similar to that of iv cyclosporine reported recently. CONCLUSIONS: This study suggest that CAP is an effective therapy in patients who are refractory to conventional medications including iv corticosteroid. Increased remission rates should be expected in refractory patients with moderately severe UC.
Cohort Studies ; Colectomy ; Colitis, Ulcerative ; Cyclosporine ; Cytapheresis ; Humans ; Inflammatory Bowel Diseases ; Leukapheresis ; Recurrence ; Retrospective Studies ; Ulcer

BACKGROUND: The Sysmex SE-9000 and R- 3000 automated cell counters provide estimates of immature cells referred to as immature myeloid information (IMI), hematopoietic progenitor cells (HPC), immature reticulocyte fraction (IRF) as high and medium fluorescent reticulocytes, and high fluorescence ratio (HFR) as high fluorescent reticulocytes. The aim of this study was to evaluate whether these parameters were useful to refine apheresis timing of peripheral blood stem cell (PBSC) harvest. METHOD: For 140 peripheral blood harvest procedures of 26 patients, pre-harvest peripheral blood (PB) WBC, mononuclear cells (MNC), IMI, HPC, CD34-positive cells, reticulocyte (%, number), IRF and HFR were tested and compared with harvested CD34-positive cell content. RESULTS: Correlation coefficients between pre-harvest WBC, MNC, IMI, HPC, CD34-positive cells, reticulocyte %, reticulocyte number, IRF and HFR of PB and harvested CD34-positive cell content were 0.15, 0.06, 0.60, 0.78, 0.77, 0.004, 0.06, 0.28 and 0.40. Applying the criteria IMI 300X10degrees/L, HPC 5X10degrees/L and CD34-positive cells 5X10derees/L of PB on the first day of 30 cycles of harvests, positive predictive value to predict the mean CD34+ cell count over 0.5X106/kg per one leukapheresis and negative predictive value to predict the mean CD34+ cell count less than 0.5X10derees/kg per one leukapheresis were 73.3%/93.3%, 57.8%/90.9% and 78.6%/ 93.7% respectively. CONCLUSION: Pre-harvest PB IMI and HPC of Sysmex SE-9000 are comparable with PB CD34- positive cells in terms of correlation with harvested CD34-positive cell content. For PB IMI and HPC are simple, inexpensive and rapid to get results, PB IMI and HPC are useful to refine apheresis timing of PBSC harvests and to screen poor-mobilizers.
Blood Component Removal* ; Cell Count ; Fluorescence ; Hematopoietic Stem Cells ; Humans ; Leukapheresis ; Reticulocyte Count ; Reticulocytes ; Stem Cells*