Summary:megalovirus, as well as a low incidence of graft-versushost disease. 3,8,9 The major problem of long-term UCB banking is the One of the main problems for the establishment of umbilical cord blood (UCB) banks is the storage space storage space required for unprocessed units. Several attempts have been made to reduce the UCB volume withneeded for the frozen samples. The aim of this study was to find a method of reducing the volume of UCB out loss of progenitor cells. + + + cell quantification were done in whole blood and in the isorecovery of progenitor cells using rouleaux formation induced by 6% hydroxyethyl starch and centrifugation to lated fractions. The average volume of the 19 UCB samples processed was 103 ml. Separation by centrifugreduce both erythrocytes and plasma. Most of these techniques imply the addition of exogenous materials and are ation led to a mean volume reduction of 56% with red cell depletion of 59%. The white blood cell recovery was carried out in an open system with the risk of contamination. of 72% with a significant CD34 + + + cell recovery of 87%. This seems a promising method for cord blood volumeThe aim of this study was to find a method for volume reduction of UCB by centrifugation with partial removal of reduction and enrichment of CD34 + + + cells. Keywords: cord blood processing; CD34 + cells; cord red cells and plasma without significant losses of the HPC CD34 + cells, using a closed system for collection and problood bank cessing.Bone marrow transplantation plays an important role in the Materials and methods treatment of many congenital and acquired haematologic diseases, either malignant or benign.1 The early haematopoietic progenitor cells are essential for haematologic and UCB collection bags immunologic bone marrow reconstitution and express the A triple blood collection system (R1656B; Baxter, Lisbon, surface glycoprotein CD34.2 It is generally known that Portugal) with a 450 ml bag containing citrate-phosphatehuman umbilical cord blood (UCB) is a rich source of dextrose-adenin (CPD-A) anticoagulant was used to collect haematopoietic progenitor cells (HPC-CD34 + ) and has been the UCB. The volume of CPD-A was reduced to 20 ml successfully used as a source of progenitor cells for bone transferring the excess to the SAG manitol satellite bag, marrow reconstitution. [3][4][5][6][7] which was then sealed and removed, keeping the system Until 1996 more than 100 cord blood transplants had closed. been performed in children or adults with total or partially HLA-matched donors. 8 The UCB offers several advantages over bone marrow and peripheral blood as it is available UCB collection without risk to mother or infant, easy and cheap to collect, and has decreased transmission of infection, namely cyto-UCB were collected from full-term pregnancies with informed consent from the donors following delivery and clamping of umbilical cord. UCB was collected from the umbili- were stored at 4°C and processed within the following 24 h.

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Monitorisation of PB haematopoietic cell count has been used as a reliable predictor of peripheral blood progenitor cell (PBPC) yields in healthy donors and patients following 5 day G-CSF mobilization. Routinely this evaluationn is performed by CD34+ cell count using flow cytometry and more recently by haematopoietic progenitor cell (HPC) count in a hematologic cell analyser. The aim of the present ongoing study is the comparison of PB CD34+ cell counts vs HPC in predicting the apheresis yield both for adult healthy donors and patients. From January to the end of July 2007, 21 healthy donors and 30 patients with hematologic malignancies (NHL=16, MM=11, HD=3) underwent G-CSF mobilization. On the 5th day of mobilization WBC and HPC counts were assessed by the Sysmex XE-2100 analyser in EDTA PB samples. CD34+ cell enumeration was performed in the same samples by standard flow cytometric assay. The healthy donors showed a good PB mobilization with a median of 82 CD34+cells/μl (range 23–193) and 161 HPC/μl (22–548). The majority of donors collected in one apheresis sufficient cells for the allogeneic transplant (>4x106 CD34+/kg bw receptor). The total amount of cells collected per kg bw donor was 8.2x106 CD34+/kg and 21.7 x106 HPC/Kg. Our data showed that HPC counts were significantly higher than CD34+ counts (2.0 fold for PB and 2.6 fold for PBPC). In relation to the patient group they showed a wide variation of PB CD34+ cells/μl (median 9, range 2–177) as well as HPC/μl (median 42, range 0–376). Patients underwent 1 to 3 apheresis to achieve >2x106 CD34+/kg. The total amount of cells collected was 2.1x106 CD34+/kg (range 0.2 – 13.6) and 5.0 x106HPC/kg (range 0.3 – 50.1). As shown for healthy donors, patient HPC counts were significantly higher than CD34+ counts (4.6 fold for PB and 2.4 fold for PBPC). Our results showed that for both groups of individuals PB HPC counts correlated with CD34+ cell counts and these numbers predicted the PBPC yield. Thus, we set a threshold on 5th day PB counts of >10 CD34+ cells/μl or >20 HPC/μl and analysed the PBPC collections. The majority of donors (17/21) presented in the PB >10 CD34+ cells/μl and >20 HPC/μl, whereas in the patient group the majority (18/31) had >20 PB HPC/μl and only 13/31 showed >10 PBCD34+ cells/μl. With PB counts above the threshold levels both donors and patients achieved the minimum requirement for hematopoietic transplant. Patients with PB counts of <10 CD34+ cells/μl or < 20 HPC/μl had a low probability of collecting > 2x106 CD34+ cells/kg in a total of 3 apheresis. The threshold on PB counts of >10 CD34+ cells/μl or >20 HPC/μl established in the current study is similar to those reported by other groups. In summary, HPC counts can be used to decide the timing of PBPC collection, as it takes only a few minutes to be executed. At the present time HPC counts can not replace CD34+ cell enumeration, as it is not yet determined the HPC dose equivalent to the current established CD34+ cell dose necessary for transplantation. Furthermore, it remains to be determine the role of the HPC content of the graft may be important in the haematopoietic recovery of patients.