Publication only EBMT 2013 Stem cell biology R1470 Mobilization of PBSCs in heterozygous-for-β thalassemia donor by addition of plerixafor after failure of mobilization R1469 with G-CSF alone for (TcR) αβ T lymphocytes depletion in Traffi cking of CD34+ cells into the peripheral circulation haploidentical transplant in thalassemia patients during collection of stem cells by apheresis: experience of P Sodani (1), A. Lanti (2), M.L. De Simone (2), E. Fiorelli (2), a single centre K Paciaroni (1), G De angelis (1), C Alfi eri (1), A. Roveda (1), A. Isgrò (1), A. Dattola, R. Fedele, E. Massara, G. Cornelio, L. Imbalzano, G. Ressa, C Gallucci (1), M. Ribersani (1), L. Cardarelli (1), M Marziali (1), C. Garreff a, A. Meliadò, M. Cannatà, I. Callea, A. Pontari, E. Spiniello, F. Torelli (1), G. Adorno (2), O.m. Chiru (2), J Gaziev (1), G Lucarelli (1) P. Scaramozzino, D. Princi, G. Irrera (1)Mediterranean Institute of Hematology (Rome, IT); (2)Policlinico AO Bianchi Melacrino Morelli (Reggio Calabria, IT) Tor Vergata (Rome, IT) Introduction: defi nition of standardized product is a product with Hematopoietic stem cell transplantation off ers the only chance of components having safe capacity of stable hemopoiesis reconsti- cure for patients with thalassemia. Haploidentical transplantation tution (effi cacy) without unseen eff ects or reactions (safety). Main may extend this possibility to the 50% to 60% of the patients who objective of all processing facilities is to have CD34+ cell products lack a suitably matched familial donor or an HLA-identical unre- standardized. To date, we did not have a consensus to defi ne the lated donor. Family members were assessed for HLA compatibility cell product, but we tried to standardize the mobilization’s capa- by serological methods or by high-resolution molecular analysis. city in aphaeretic collection products (A-HPC). Two heterozygous for β thalassemia family donor for the very Material and Methods: we evaluate n. 580 A-HPC and related sam- young son ( 4 years old and 7 years old respectively ) candidate ples. Quantifi cation/enumeration of CD34+ in absolute count was to haplo-SCT for thalassemia major did not achieve the expected performed with ISHAGE Protocol by fl ow cytometry FACSCalibur, target >8 x10 6 CD34+/kg recipient with conventional mobiliza- Stem kit and Trucount test tubes. lion (G-CSF 10 mcg/kg daily) and were successfully rnobilized with Results: CD34+ cells number in cell products collected by aphae- plerixafor. Following informed consent for off -Iabel use, the both resis exceeds 7,6 times CD34+ cells number calculated on periph- donors received a single subcutaneous injection of 24 mgs plerix- eral venous blood’s samples before collection, with statistical afor in combination with G-CSF with no side eff ects; 12 hrs later: signifi cance 0,00 (p=0,00). in the fi rst donor we observed circulating CD34+: 105/mcL (WBC Release’s calculation of CD34+ in pre-collection samples is 96 cell/ul 76x103/mcL), CD34/Kg recipient (post TcRα β T lymphocytes selec- (range 20-1404), in A-HPC is 975 cell/ul (range 64-17101). tion 11.2x106 CD34+/Kg,). In the second donor, using the same Collection’s performance for CD34+ (cell/ul) is 84% and it is inde- mobilization strategy the circulating CD34+ was 110/mcL (WBC pendent from processing of blood, diagnosis and conditioning 59.000x103/mcL), CD34/Kg recipient (post TcRα β T lymphocytes regimen. It depends from cellularity (cell/ul) on peripheral venous selection 8,1x106 CD34+/Kg,). The median time to neutrophil blood. and platelets engraftment was at 12 days and 14days, respec- Conclusions: Collected cells are present on peripheral venous tively.Using this new strategies in both haploidentical transplant blood at the begining of A-HPC collection. All patients show in thalassemia patients we observed more rapid immunological having a signifi cant increase of CD34+. A-HPC quality is directly reconstitution after 60 days post transplant, with an increase of dependent from peripheral venous blood quality. T cells (CD4+ and CD8+), B cells (CD19+) and NK cells (especially CD3-CD16+, with cytotoxic potential) in the peripheral blood of these patients. In conclusions: For all “poor mobilizers donors, included “ heterozygous for β thalassemia donor adequate trans- plant CD34+ cells dose was obtained by combination of Plerixafor with G-CSF with no side eff ects. R1471 Autologous stem cell transplantation: analysis of haematimetric parameters R. Guerrero Camacho, M. Gasior Kabat, K. Humala Barbier, P. Gomez Prieto, R. De Paz, A. Marcos, R. Arrieta Gallastegui La Paz University Hospital (Madrid, ES) Objectives: To investigate erythrocyte parameters oriented to program and optimize the apheresis procedure based on the stem cell harvest effi ciency. Materials and Methods: Medical records of 294 patients diag- nosed with Non-Hodgkin Lymphoma (155), Hodgkin Disease (78) and Multiple Myeloma (61) were reviewed retrospectively. The apheresis procedures were conducted between 2000 and 2010. Effi ciency was calculated according to this formula: Effi ciency (%)=Obtained Mononuclear Cells (MNC)(109)/[( Total Processed Volume/Volemia) x (pre-apheresis MNC(109)+post-apheresis MNC (109)/2)]. S519 Studied parameters were: Haemoglobin (HB), Hematocrit (HT), concentration of 10% DMSO. For pediatric and clinically indicated Mean Corpuscular Volume(MCV), Mean Corpuscular Hemoglobin patients fi nal concentration is 5% DMSO. To assess the PBSCs’ (MCH), Mean Corpuscular Hemoglobin Concentration (MCHC) quality, we evaluate total nucleated cells (TNC), CD34+ content and Red Blood Cell Distribution Width (RDW). These parameters and microbiological contamination (aerobic, anaerobic and myco- were compared between a Group with Effi ciency<60% and a logical cultures); an aliquot of each bag is stored (cell viability and Control Group with Effi ciency>80% using the Non-parametric clonogenic assays). Products are frozen in a rate-controlled pro- Mann-Whitney U-Test. grammed freezer (Nicool Plus PC, Air Liquide) and stored between Results: Patients with Hodgkin Disease showed a lower Effi ciency -150 and -196ºC in vapor and/or liquid phase of liquid nitrogen. than patients with Multiple Myeloma (p=0.015). In general, the At infusion time, frozen PBSC products are thawed at 37ºC and can Mean Effi ciency value of the patients studied was between 60 be directly infused (bedside thaw) or require post-thawed proce- and 80%, only a 6,5% of the patients (n=19) showed a Mean Effi - dures. Product’s quality is indirectly evaluated by engraftment’s ciency<60% and 48% showed a Mean Effi ciency>80%. Mean MCV success (Absolute Neutrophil Count – ANC). and MCH was lower in patients with Effi ciency>60% (p=0.026). Results: In 2011, 95 autologous PBSC from 69 patients (26% Conclusion: Pre-apheresis alterations in MCV, MCH and RDW pediatric – 5 months/16 years) were processed at our laboratory. values should be considered as adverse factors that can infl uence 58 PBSC were processed on the same day (61%) and the remain- the stem cell collection procedure. ing 37 (39%) were stored overnight at 4ºC and processed in the next morning. Average product volume was 128mL; 21 products (22%) required volume reduction. PBSC’s quality results: TNC aver- age 3.34x108/Kg, CD34+ average 4.65x106/Kg and no microbio- logical contamination. 65/95 PBSCs collected (68%) were infused in 50/69 patients (72.5%); only 1/65 PBSC (1.5%) required post- thawing procedure (DMSO removal). 49 /50 patients (98%) recov- ered ANC between d+6 and d+17 (average 11.51 days). Conclusions: Results confi rm the quality of the new PBCS cryo- preservation protocol. R1473 Clinical and haematological features of oncohaematological patients with low content of CD34+ cells in autotransplant S. Gritsaev, A. Kuzajeva, I. Zapreeva, Z. Chubukina, S. Tiranova, V. Balashova, A. Seltcer, K. Abdulkadirov Russian Institute of Hematology (St.Petersburg, RU) There are some conditions to get succesful engraftment after autologous stem cell transplantation (AutoSCT)and the main one is the number of CD34+ cells in the autotransplant which has to be ≥2x106/kg. The aim of the study was to characterize the patients with the autotransplant failure. There were 14 patients whose autotransplant contain less than 2x106 CD34+ cells/kg. For the control group 36 patients have been chosen. Leukoapheresis has been done with COBE Cpectra (version 6.1, Gambro). Median age of the patients was 47.5 years (19-62). There were next diagnoses: 6 patients with myeloma, 6 with nonHodgkin lymphoma and 2 with acute lymphoblastic leuke- mia. Before autologous stem cell transplantation (AutoSCT) the patients were treated with 1-4 regimens of chemotherapy. The mobilisation of stem cells was performed with 3 regimens. So the patients were divided into the 3 groups. In the fi rst group 5 patients were treated with G-CSF only. In the second group 8 patients were treated with Cph 1-3 g/m2 plus G-CSF. In the third group 1 patient was treated with high dose chemo- therapy plus G-CSF. The median number of mononuclear cells R1472 in the autotransplants was 1.9x108/kg (1.0-5.8). The median Peripheral blood stem cell cryopreservation: a safety and number of CD34+ cells in the autotransplants was 0.51x106/kg quality analysis (0.03-0.72). C. Espadinha, A. Barbosa, M. J. Gutierrez, E. Pereira, M. Abecasis There was no any diff erence between study and control groups in IPO Lisbon (Lisbon, PT) the age, number of previous chemotherapy, total dose of antra- cyclines and number of aphereses. At the same time there was Introduction: Autologous peripheral blood stem cells (PBSC) association between the low number of CD34+ cells in the auto- transplantation relies on eff ective preservation of those cells. Cell transplant and the dose of Cph less than 2.5 g/m2.