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Viability of non cryopreserved CD34+Stem Cells . On behalf of the « Western Algerian Group of Bone Marrow Transplant » WAG-BMT Haematology and Cell Therapy Depatment Hemobiology Department. Introduction.
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Viability of non cryopreserved CD34+Stem Cells On behalf of the « Western Algerian Group of Bone Marrow Transplant » WAG-BMT Haematology and Cell Therapy Depatment Hemobiology Department
Introduction • Autologous stem cell transplantation (ASCT) after a high dose conditioning is an established treatment modality with definitive indications for many hematological disorders. • However, this line of treatment requires many expensive resources, such as freezing of the harvest product in order to maintain cell viability until stem-cell reinfusion and the use of growth factors for the management of neutropenia.
Aim of the study • To study the viability of CD34+ stem cells stored at +4°C • To demonstrate the feasibility and the safety of the ASCT without any cryopreservation neither the use of growth factors.
Study design • STEP 1: Short Conditioning regiments ASCT in MM (MEL200) in 75 patients ASCT in HL (CBV) in 17 patients • STEP 2: Long conditioning regiments ASCT in HL (BEAM) in 02 patients ASCT in HL and NHL (EAM) in 15 patients
STEP 1Material and methods 75 patients with Multiple Myeloma Figure 1. Schema of procedure.
STEP 1Material and methods 17 patients with Hodgkin Lymphoma
STEP 1Material and methods Wannesson L et al. Annals of Oncology 18: 623–632, 2007
STEP 1Material and methods • A 2ml sample was taken from 10 collection bags and then was sent to the flow cytometry laboratory where it was stored in the refrigerator at +4°C. • A CD34+ stem cells count were performed using a “Lyse no wash double platform CD34+ flow cytometry assay” • The viability of CD34+ cells was assessed with 7'AAD immediately after the end of apherisis (H0) then at (H24), (H48) and finally at (H72).
STEP 1Results y = -1,06x + 99,3 R2 = 0,9571 p=0.05 CD34+ stem cells viability kenetic curve
STEP 1Results Brahimi M et al. Revue Algerienne D’hematologie 2011 ; 5 :46-48.
STEP 1Results n=200 Time of storage of the Harvest bags at +4°C
Mel200 protocol Bekadja MA et al. Hematol Oncol Stem Cell Ther 2012; 5(1): 49-53 Bekadja MA et al. EBMT 38th meetting: Geneva, Switzerland 1-4 april 2012.page 140 (Abstract 905)
CBV protocol Bekadja MA et al. Revue Algerienne D’hematologie 2011 ; 5 :50-53.
STEP2Material and methods 15 Patients with Hodgkin Lymphoma Bekadja MA et al. EBMT 38th meetting: Geneva, Switzerland 1-4 april 2012.page 162 (Abstract 1140) Talhi S et al. Hématologie, Vol. 1, Supplement 1, Mars 2012: pp 213 (Abstract)
STEP2RESULTS n=30 Time of storage of the Harvest bags
STEP2RESULTS Brahimi M et al. Unpublished data.
Conclusion • We conclude that high-dose chemotherapy with non-frozen peripheral stem cells is safe in terms of haematopoietic reconstitution even without using growth factors.
Department of Hemobiology EHU 1st November, Oran, Algeria
Department of Hematology and Stem Cell Therapy EHU 1st November Oran, Algeria
Chief of Biology Pôle in EHU 1stNovember Professor Mohamed Amine BEKADJA Bekadja MA et al. Revue Algérienne d’Hématologie 2011 ; 5 :50-53. Bekadja MA et al. Hematol Oncol Stem Cell Ther 2012; 5(1): 49-53 Bekadja MA et al. EBMT 38th meetting: Geneva, Switzerland 1-4 april 2012., (Abstract 905) Bekadja MA et al. EBMT 38th meetting: Geneva, Switzerland 1-4 april 2012, (Abstract 1140)