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C ollection h a ematopoietic stem cells by leukapheresis - the experience of BTC in Ljubljana.

C ollection h a ematopoietic stem cells by leukapheresis - the experience of BTC in Ljubljana. Dragoslav Domanovi ć,MD,MSc Zavod Republike Slovenije za transfuzijsko medicino, Šlajmerjeva 6, 1000 Ljubljana, Slovenija. History of haematopoietic stem cell (HSC) collection & transplantation.

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C ollection h a ematopoietic stem cells by leukapheresis - the experience of BTC in Ljubljana.

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  1. Collection haematopoietic stem cells by leukapheresis- the experience of BTC in Ljubljana. Dragoslav Domanović,MD,MSc Zavod Republike Slovenije za transfuzijsko medicino, Šlajmerjeva 6, 1000 Ljubljana, Slovenija

  2. History of haematopoietic stem cell (HSC) collection & transplantation • 1960 • attempt of allogeneic bone marrow (BM) transplantation in Maribor • 1988 • collection of autologous BM • 1989 • successfull transplantation of auto- & allo- BM • 1994 • collection of auto- & allo- peripheral blood stem cells (PBSC) – CS 3000; in 1998 Amicus • 2000 • immunomagnetic isolation of CD34+ cells

  3. Annual number of PBSC collections

  4. Study design • Retrospective analysis • Predictive value of CD34+ cells in blood of donors before collection for achieving a yield of > 2x106 CD34+ cells /kg of recipient’s BW

  5. Material and methods • N = 74 leukafereses • 54 cases of PBSC collections from January 2002 to July 2003 • 39 patients (autologous collections) • 15 donors (allogeneic collections) • Target number of collected cells • > 2 x 106 celic CD34+ /kg of recipient’s BW • Mobilisation: rH G-CSF filgrastim (Neupogen) • 10 μg/kg BW s.c. in two divided doses daily • patients: G-CSF or G-CSF + chemotherapy • donors: G-CSF

  6. PBSC collection using cell separator • Standard volume leukapfereses (SVL) (processing of the 2 blood volumes) • Cell separator Amicus (Baxter Healthcare, IL, USA) • anticoagulans ACD and blood ratio 1 : 12. • Blood volume processed in one cycle was determined according to the number of WBC in donors blood

  7. Laboratory procedures • Blood cell counts • Electronic counter CellDyne 3200 (Abbott Laboratories, IL, USA) • CD34+ cells • flow cytometer Coulter Epics XL – MCL (Beckman–Coulter/Immunotech, FL, USA) ISHAGE protocol • Collection efficiency (%) • calculated

  8. Statistical analysis • Pearson’s correlation coeficient • Multiple linear regresion • Mann -Whitney U test • Receiveroperating characteristics analysis (ROC)

  9. Auto- Allo- Total Number of aphereses (N tested) 58 16 74 Number of cases 39 15 54 Females 12 8 20 Males 27 7 34 Average age 47 34 44 Averagenumber of apher./case 1.48 1.06 1.37 1 apheresis/case 22 (56%) 13(87%) 35 (65%) 2 aphereses/case 16 (41%) 1 (13%) 17 (33%) 3 aphereses/case 1 (3%) 0 1 (2%) Demographics

  10. Results - Blood Cell Counts * p<0.05

  11. Collection efficiency

  12. Relation between the mobilized & collected CD34+ cells 16.00 r = 0,82 14.00 p < 0,05 12.00 10.00 number of CD34+ cellsx10e6/kg /body weight 8.00 6.00 4.00 2.00 . 0 00 140 0 20 40 60 80 100 120 160 180 200 220 240 260 280 number of CD34+ cells x10e6/l

  13. Cut-off value of CD34+ cells before collection CD34_ 42 x 10e6/l cut – off value 100 Sensitivity = 92% Specificity = 87 % 80 60 Sensitivity 40 20 ROC curve 0 0 20 40 60 80 100 100-Specificity

  14. Possibility to collect >2x10e6 CD34+ cells/kg BW

  15. Troubleshootings with Amicus device Total number of troubleshootings: 31/1094 (2,83%)

  16. Donor adverse reactions Mild type of reactions in total 11/74 (14%) 4 (36%) 2 (18%) 5 (46%) Number of collections

  17. Conclusion I • On the Amicus cell separartor using SVL we can collect yield target of > 2 x 106 CD34+ cells/kg body weight of recipient with 1,48 collections per case in average. • Collection efficiency of MNC is 61% and of CD34+ cells is 55% • Low platelet loss is related to the specific separation method employed using Amicus.

  18. Conclusion II • The number of CD34+ cells in donor’s blood can be used for prediction of CD34+ cells yield and therefore for the optimal time for the beginning of the collection. • The cut-off value ≥42 x 106/l CD34+ cells in blood before collection indicate a high probability to collect > 2 x 106 CD34+ cells/kg body weight of recipient.

  19. Conclusion III • Collection of PBSC using Amicus cell separator is safe and efficatious with acceptable number of troubleshootings.

  20. Sources of Blood Stem Cells for Transplantation • Bone Marrow • Peripheral Blood • Cord Blood

  21. Peripheral Blood Stem Cells • Advantages - disadvantages • Mobilisation with G - CSF • Collection of MNC by leukafereses

  22. Mobilisation of PBSC with G-CSF • G-CSF • G-CSF + chemotherapy • Chemotherapy

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