Peripheral blood stem cell collection with a blood cell separator

Forty-three patients with malignant nonmyeloid diseases underwent peripheral blood stem cell collections on an apheresis system (Spectra, COBE BCT, Lakewood, CO). Collections took place during the white cell (WBC) recovery phase following conditioning chemotherapy. One hundred two procedures were done after chemotherapy alone, and 72 procedures after chemotherapy plus granulocyte-colony-stimulating factor (G-CSF). Four centrifugal separation factors were tested. One and one-half patient blood volumes were processed in each procedure. The mean volume of the collected component was 158 +/− 16 mL. After chemotherapy alone, the procedures provided a mean of 0.8 × 10(8) WBCs per kg and 2.3 × 10(4) colony-forming units-granulocyte macrophage (CFU-GM) per kg of recipient body weight. The mononuclear cell percentage in the components increased with the centrifugal separation factor from 85 to 96 percent. In parallel, platelet contamination increased from 2.1 to 3.8 × 10(11). The collect hematocrit ranged from 1.0 to 2.5 percent (0.01-0.025). The collection efficiency for mononuclear cells and CFU- GM also increased with the centrifugal separation factors from 52 to 70 percent for mononuclear cells and from 55 to 68 percent for CFU-GM. Collections performed after G-CSF-stimulated mobilization were characterized by a higher neutrophil contamination independent of centrifugal separation factor, which gave a mean mononuclear cell percentage of 64 percent in the collected component. The average yield for these procedures was 2 × 10(8) WBCs per kg and 28 × 10(4) CFU-GM per kg.(ABSTRACT TRUNCATED AT 250 WORDS)     

Plateletapheresis with a next generation blood cell separator

Procedure time has been identified as the most important element in apheresis platelet donor retention. Fenwal has developed a next generation apheresis system, the Amicus, with the intent of efficiently producing high platelet yields with low WBC content in much shorter processing times than currently available. This report describes the Amicus and presents results of a clinical trial of an Amicus prototype and a comparison to Fenwal CS 3000+. Thirty Amicus double-needle plateletapheresis procedures were evaluated. Average processing time was 61 ± 16 min with 63% of the processing times ≤ 60 min. The average preplatelet count was 246 ± 46 × 10³/μl, platelet yield 4.4 ± 1.2 × 10¹¹ plt, collection efficiency 73 ± 14%, platelet collection rate 0.075 ± 0.016 plt × 10¹¹/min and citrate toxicity incidence 3%. A comparison of double-needle procedures, 20 Amicus and 20 CS 3000+, showed that Amicus had significantly shorter (P < .05) processing times (64 ± 17 vs. 86 ± 10 min) and higher (P < .05) platelet collection rates (0.070 ± 0.017 vs. 0.054 ± 0.018 plt × 10¹¹/min) but comparable (P < .05) platelet yields (4.4 ± 1.4 vs. 4.7 ± 1.0 × 10¹¹ plt). Comparison of 13 single and 13 double-needle Amicus procedures showed comparable (P < .05) processing times (71 ± 13 vs. 65 ± 18 min), platelet yields (4.7 ± 1.0 vs. 4.6 ± 1.5 × 10¹¹ plt), collection efficiency (74 ± 7 vs 74 ± 17%), and platelet collection rates (0.068 ± 0.020 vs 0.072 ± 0.018 plt × 10¹¹/min). Using normal probability plots of WBC content at 95% confidence level, Amicus can provide products with <5.0 × 10⁶ WBC or <1.0 × 10⁶ WBC in 99.7% or 92.7% of collections, respectively, compared with 69.0% or 44.0%, respectively, for CS 3000+. We found Amicus was able to provide equivalent quantities of platelets with less WBC content in significantly shorter processing times than CS3000+ as well as shown encouraging results for single-needle procedures. J. Clin. Apheresis 12:55–62, 1997. © 1997 Wiley-Liss, Inc.     

Exchange transfusion in sickle cell disease using a continuous-flow blood cell separator

An exchange transfusion was performed preoperatively on a patient with sickle cell disease using a continuous-flow blood cell separator. An exchange of 2,825 ml red blood cells achieved a hemoglobin A level of 90.8 per cent. The continuous-flow blood cell separator appears to offer a safe and effective method of exchange transfusion in sickling disorders.     

5-day storage of platelets collected on a blood cell separator

One of the earliest devices available for plateletpheresis is the Haemonetics system; this machine has been updated recently to permit the use of software in a closed system and thus storage of the collected platelets beyond 24 hours. The authors examined the in vitro and in vivo function of platelets collected on the Haemonetics AutoSurge machine and stored for 5 days in two separate 1000-mL CLX bags. The average count per bag was 1.8 +/- 0.4 x 10(11) in approximately 200 mL of plasma. Immediately following collection, the platelet response to ADP and epinephrine represented 78 and 35 percent, respectively, of the preapheresis values. Aggregation to single stimuli subsequently decreased to 29 and 0 percent, respectively by Day 5. This response is equal to or better than the response reported with the Fenwal CS-3000, the only other plateletpheresis device routinely used for long-term storage. The pH of the preparations was well maintained throughout storage, and there was little alteration in hypotonic shock response or serotonin uptake. Serotonin release decreased consistently. The morphology scores indicated good maintenance of shape immediately following collection; this subsequently decreased after 5 days of storage. Bacterial cultures were negative in all instances. The 51Cr in vivo survival and recovery was good with 65.5 +/- 7.1 percent recovery and an average survival of 7.3 +/- 1.3 days (multiple hit; n = 5). The data indicate that storage of the Haemonetics plateletpheresis product is feasible and that the product is as good as others currently available.     

Extended storage of single-donor platelet concentrate collected by a blood cell separator

Use of a sealless blood pathway in a blood cell separator (CS-3000, Fenwal) permits collection of platelets in a "closed system" when saline and anticoagulant solutions are integrally attached; this in turn allows storage of instrument-collected platelet concentrates (PCs) beyond 24 hours. To evaluate extended storage of high yield PCs, cells collected with the instrument were stored (200 ml plasma) for 8 days (flatbed agitation) in either 3-liter polyvinylchloride (PL 146) containers (n = 6), polyolefin bags (PL 732) (n = 8), or two 1-liter polyolefin (double PL 732) containers (n = 8). A mean of 4.45, 4.09, and 3.94 X 10(11) platelets were stored in PL 146, single PL 732, and double PL 732, respectively; total white cells per container averaged 0.3, 0.2, and 0.2 X 10(9) for the three container systems. By day 1, platelet pO2 dropped to 14 and 16 torr in PL 146 and PL 732 PCs (pCO2, 127, and 82 torr). In contrast, double PL 732 maintained high pO2 (approximately equal to 80 torr) and low pCO2 (approximately equal to 30 torr) through day eight. Glucose declined at faster rates in PL 146 and single PL 732 containers, while lactate increased more rapidly (338 and 197 mg/dl of lactate on day four vs. 116 mg/dl for double PL 732 units). Morphology scores dropped from 400 to 98 (PL 146) and 216 (PL 732) at day four (pH values of 6.3 and 7.0), while a score of 330 was seen in double PL 732 PCs.(ABSTRACT TRUNCATED AT 250 WORDS)     

5-day storage of single-donor platelets obtained using a blood cell separator

Platelets were collected from normal donors via a blood cell separator (Fenwal CS-3000). Platelets were stored initially in two separate 1000-ml bags (average count per bag, 2.3 +/- 0.5 x 10(11)) in 100 ml of autologous plasma for 5 days. Little change in platelet count was noted after 5 days of storage; however, the white cell count fell from 5.1 +/- 1.7 x 10(9) at Time 0 to 3.4 +/- 2.3 x 10(9) per l at Day five. The initial lactate values were 32 +/- 11 mg per dl and rose to 165 +/- 28 mg per/dl by 5 days. Platelet aggregation was impaired both by the collection procedure and during storage: whereas the response to ADP of the donors' platelets before the procedure was 100 percent, samples taken from the product immediately after collection had only a 45 percent response, which fell to 12 percent by Day 5. Aggregation using epinephrine was similarly affected, with a 75 percent response after collection and 0 percent response by Day 5. The plasma beta-thromboglobulin (beta-TG) level was high, both after collection (5.0 micrograms/ml at Time 0) and after storage (11.0 micrograms/ml), indicating a considerable effect of collection on platelet alpha granule release. In vivo recovery of these platelets was very good at 67 +/- 6 percent, with an average survival of 7.3 +/- 1.4 days (multiple hit; n = 4).(ABSTRACT TRUNCATED AT 250 WORDS)     

Concentration of bone marrow mononuclear cells using a programmable blood cell separator.

Bone marrow was collected from adult patients with various solid tumors who consented to participate in a study of myelo-ablative chemotherapy followed by autologous bone marrow rescue. Twenty marrow suspensions were processed by using standard Procedure 3 (PRO-3) for lymphocytapheresis without modification. A modified Procedure 1 (M-PRO-1) for plateletpheresis was employed for processing 34 marrow suspensions. For PRO-3, mononuclear cell (MNC) recovery was 68 +/- 22% of the starting marrow suspension (baseline), in a concentrate volume of 234 +/- 53 ml. MNC represented 59 +/- 27% of the total WBC count of the concentrate. The residual volume of RBC was 49 +/- 47 ml. For M-PRO-1, MNC recovery was 63 +/- 22% of the baseline in a concentrate volume of 200 +/- 8 ml. MNC comprised 94 +/- 7% of the total WBC count of the concentrate. RBC contamination was 7 +/- 3 ml. Hematopoietic recovery, defined as the post-transplant days when a sustained granulocyte count of 500/microL and a platelet count of 50,000/microL were achieved, was similar in the two groups and comparable to other reports utilizing other methods and equipment for bone marrow concentration. Personnel time was significantly reduced compared to other procedures for bone marrow concentration due to increased automation. Although there was no significant difference in MNC recovery between the two groups (P greater than 0.5), M-PRO-1 was clearly superior to PRO-3 because of the consistently high degree of purity of the MNC in the concentrate and minimal RBC contamination.(ABSTRACT TRUNCATED AT 250 WORDS)     

应用间断流动式血细胞分离机采集外周血造血干细胞的效果分析

目的:为寻找采集外周血造血干细胞更合适的手段,探讨间断流动式血细胞分离机采集外周血造血干细胞的效果。方法:选择中山大学第五附属医院血液风湿科2004-2006年12例行外周血造血干细胞移植住院患者。①实验对象:供者5例,男1例,女4例,一般状况良好,与患者关系为同胞妹妹3例,同胞弟弟1例,同胞姐姐1例;患者7例,男3例,女4例,23～62岁,7例为自体移植,5例为异基因移植。血液系统疾病患者9例(非霍奇淋巴瘤3例,急性淋巴细胞白血病2例,急性髓细胞白血病2例,慢性粒细胞白血病1例,骨髓增生异常综合征1例);自身免疫性疾病3例(重型系统性红斑狼疮2例,难治复发性类风湿关节炎合并干燥综合征1例)。②实验过程:每位供/患者均知情同意并签署知情同意书。对于自体患者,根据患者的疾病类型采取不同的干细胞动员化疗方案,联合化疗后7～10d,待白细胞下降至最低点,一般为≤1.0×109 L-1时,给予粒细胞集落刺激因子5μg/kg皮下注射,白细胞升至3.0×109 L-1时开始采集;对于allo-PBSCT供者直接给予粒细胞集落刺激因子5μg/kg,皮下注射,1次/d,共5d,同时应用流式细胞仪检测CD34 细胞数,至白细胞升至≥20.0×109 L-1或当CD34 细胞升高>20个/μL时采用增强型多功能血细胞分离机PBSC程序卡采集健康供者和患者外周血造血干细胞。③实验评估:分析采集效率、血液学参数、不良事件发生率、以及供、受者ABO血型不合的患者回输干细胞溶血反应等情况。结果:12例供者、患者均进入结果分析。①共进行了24次采集,其中1次采集1例,2次采集7例,3次采集3例,平均循环次数(25±5)次,采集时间(228±32)min,处理血量(7234±1205)mL,复方枸橼酸钠溶液用量为(623±96)mL,干细胞收集量为(102±21)mL,CD34 细胞采集效率为(54.3±30.7)%,细胞计数示白细胞为(156±34)×109 L-1,单个核细胞为(79.7±13.2)×109 L-1,流式细胞仪检测CD34 细胞为(10.30±4.38)×106/kg受者体质量。②不良反应轻微,24次采集过程中出现不良反应6次均为枸橼酸盐所致低钙血症反应。血红蛋白和血小板与采集前相比分别下降9.36%和11.10%。供、受者ABO血型不合的3例患者在输注造血干细胞悬液后均未出现溶血反应。③12例均获造血功能重建,无移植相关死亡。结论:用间断流动式血细胞分离机采集外周血造血干细胞效果良好,不良反应轻微,能有效的减少被采集者红细胞、血小板的丢失,对供、受者ABO血型不合者不需去除造血干细胞悬液中的红细胞,值得临床应用。     

Plateletpheresis with the new Fresenius AS 104 blood cell separator

Among the many blood cell separators introduced into the international market in these last few years, the Fresenius AS 104 represents an advanced and safe thrombocytapheresis machine whose development took advantage of extensive worldwide experience with blood cell separation. Nonetheless the AS 104 has generated most interest in West Germany and most, if not all, the studies published on its platelet collection efficiency have been carried out in that country. It is normally reported that from 2.7 to 3.5 x 10(11) platelets can be collected in approximately 80 minutes. Since these results could not be duplicated routinely in our hemapheresis unit, we set up a study by modifying the standard procedure. It was possible to reduce the procedure time and to collect platelet concentrates containing more than 4 x 10(11) cells on a routine basis by using the following procedure: ACD-A/blood ratio 1:10; Interface position 6:2; blood flow rate always exceeding 65 mL/min; rpm 1750; cell collection from 4 to 7 mL/min; volume of blood processed 3.6 L followed by the rinsing of the system with 200 ml of saline; extraction of the content of the secondary separation chamber by the action of the plasma pump working at 20 mL/min for 2 min. With this procedure the platelet yield in 34 collections exceeded 3.1 x 10(11) and averaged 4.06 x 10(11). The procedure time was reduced to 56.5 minutes with a mean blood flow rate of 62.3 mL/min. The leukocyte and erythrocyte contamination of the products were in the range of 1 x 10(7) and 1 x 10(8) respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Leukocyte transfusion and the development of the continuous-flow blood cell separator

The treatment of anemia and thrombocytopenia with allogeneic cell transfusions is an effective and well-developed technology. However, leukocyte replacement transfusion has been frustrated by the physiology of the leukocytes. To achieve effective leukocyte replacement, the continuous-flow centrifugal blood cell separator was developed, and it soon proved to be an important instrument for separation, collection, and transfusion of all the components of the blood. Thus, the continuous-flow centrifugal blood cell separator has become an important instrument in the science of blood collection and transfusion.     

Evaluation of a new protocol for peripheral blood stem cell collection with the Fresenius AS 104 cell separator

In this report we analyzed sixty leukapheresis procedures on 35 patients with a new protocol for the Fresenius AS 104. Yields and efficiencies for MNC, CD 34+ cells, and CFU-GM indicate that the new protocol is able to collect large quantities of hemopoietic progenitors. Procedures were performed processing 8.69 ± 2.8 liters of whole blood per apheresis and modifying 3 parameters: spillover-volume 7 ml, buffy-coat volume 11.5 ml, centrifuge speed 1,500 rpm; blood flow rate was 50 ml/min and the anticoagulant ratio was 1:12. No side effects were observed during apheresis procedures except for transient paresthesia episodes promptly resolved with the administration of calcium gluconate. Yields show a high capacity of the new program to collect on average MNC 17.28 ± 10.85 × 10⁹, CD 34+ 471 ± 553.5 × 10⁶ and CFU-GM 1278.7 ± 1346.3 × 10⁴ per procedure. Separator collection efficiency on average was 49.91 ± 23.28% for MNC, 55.1 ± 35.66% for CFU-GM, and 62.97 ± 23.09% for CD 34+ cells. Particularly interesting are results for MNC yields and CD 34+ efficiency; these results make the new program advantageous or similar to the most progressive blood cell separators and capable to collect a sufficient number of progenitor cells for a graft with a mean of 1.80 ± 0.98 procedures per patient. J. Clin. Apheresis 12:82–86, 1997. © 1997 Wiley-Liss, Inc.     

A technique to reduce lymphocyte contamination of plateletapheresis products collected with a centrifugal blood cell separator

It is desirable to minimize the contamination of plateletapheresis products by lymphocytes because of the role these cells may play in febrile nonhemolytic transfusion reactions, alloimmunization, platelet deterioration during storage and also to reduce donor lymphocyte loss. This study attempted to determine whether lymphocyte contamination could be reduced by rapidly depleting plasma from the separation chamber at selected intervals during plateletapheresis with a blood cell separator (CS-3000, Fenwal). Donors who provided platelet components with more than 0.10 X 10(9) lymphocytes without rapid depletion underwent a second plateletapheresis procedure in which the rapid depletion technique was used. The plasma pump was reprogrammed to remove rapidly 12 ml of plasma from the separation chamber (pump speed 48 ml/min) immediately after the plasma pump reversal. All other collection techniques were identical for all procedures. Significantly fewer lymphocytes were collected in those procedures in which rapid depletion was used (mean, 0.187 X 10(9] than when it was not used (mean, 0.331 X 10(9] (p = 0.001, n = 30). There was no effect on platelet yield, efficiency of platelet collection, processing time, or total collection time. This procedure provides a product that should be considered for routine use.     

Collection of WBC-reduced single-donor PLT concentrates with a new blood cell separator: results of a multicenter study

BACKGROUND: A new cell separator (COM.TEC, Fresenius) was recently developed aimed at efficient collection of WBC-reduced single-donor PLT concentrates (SDPs). STUDY DESIGN AND METHODS: Five German centers collected 554 WBC-reduced SDPs with help of the COM.TEC cell separator. Two multicenter cell counting studies were performed at the beginning and at the end of the study to document uniform counting results among the participating centers. RESULTS: A total of 441 (79.6%) PLT collections were included in the study according to the protocol. A total of 342 single-dose and 99 double-dose SDPs were collected. For single-dose SDPs, an average blood volume of 2826 +/- 409 mL was processed in a donation time of 55 +/- 11 minutes. Mean PLT yield of these products was 3.11 x 1011+/- 0.40 x 1011 and the WBC contamination was 0.11 x 106+/- 0.20 x 106. For double-dose SDPs (PLT count, 5.29 +/- 0.93 x 1011), 3943 +/- 639 mL was processed. The average difference between the target and the collected PLT concentration was -2.8 +/- 12.0 percent for single-dose SDPs and -1.8 +/- 9.5 for double-dose SDPs, respectively. The collection efficiency was 53.7 +/- 5.8 percent for single-dose SDPs and 58.2 +/- 6.2 percent for double-dose SDPs. If all results of each sample from the counting study were set to unity (to the mean over all centers), most PLT determinations were very similar to the mean, for example, near or 1 if set to unity. CONCLUSION: The COM.TEC machine makes it possible to obtain WBC-reduced SDPs that comply with current standards.     

Application of a new LDL apheresis system using two dextran sulfate cellulose columns in combination with an automatic column-regenerating unit and a blood cell separator

Extracorporeal procedures for selective removal of low-density lipoproteins have become a promising new approach for treatment of severe familial hypercholesterolemia. We tested efficacy and safety of a new LDL apheresis system by using two dextran sulfate cellulose adsorbents (Liposorber LA 15TM from Kanegafuchi) under the control of an automatic column-regenerating unit for continuous alternate adsorption and desorption. Plasma was taken from a continuous-flow blood cell separator (model IBM/Cobe 2997) allowing an extracorporeal circuit from one cubital vein to another. A 57-year-old male with drug-resistant heterozygous familial hypercholesterolemia accompanied by moderate hypertriglyceridemia and severe coronary artery disease has been treated every 2 weeks for 3 months so far. Treatment of 4-5 liters of plasma resulted in a mean decrease of total cholesterol from 355 to 111 mg/dl (9.20 to 2.88 mmol/l), of LDL cholesterol from 272 to 49 mg/dl (7.05 to 1.53 mmol/l), and of apolipoprotein B from 175 to 44 mg/dl. HDL cholesterol, apolipoprotein A-I, and other plasma proteins did not substantially change apart from hemodilution. No side effects were seen. This new technique of LDL apheresis represents a very effective and safe method for treatment of drug-resistant familial hypercholesterolemia without or with concomitant hypertriglyceridemia.