Automated washing of autologous haematopoietic stem cell grafts

Authors: L. Adamusová 1,2;  Z. Kořístek 1,3;  J. Smejkalová 1;  M. Navrátil 1,3;  L. Grebeníček 1;  I. Tvrdá 1;  M. Michaliková 1;  R. Hájek 1,3
Authors‘ workplace: Klinika hematoonkologie, Fakultní nemocnice Ostrava 1;  Katedra biologie a ekologie, Přírodovědecká fakulta, Ostravská univerzita 2;  Lékařská fakulta, Ostravská univerzita 3
Published in: Transfuze Hematol. dnes,24, 2018, No. 2, p. 115-122.


Introduction: High-dose chemotherapy with autologous transplantation of haematopoietic stem cells represents a standard treatment of various haematological malignancies. Prerequisites for successful transplantation include collection of a sufficient number of haematopoietic stem cells, their processing, cryopreservation, and storage guaranteeing their complete viability until transplant. To maintain viability of haematopoietic stem cells, it is necessary to use a cryoprotective agent for cryopreservation. Dimethyl sulfoxide is the most commonly used cryoprotective agent, typically at a concentration of 10% in the grafts. Reinfusion of hematopoietic stem cells containing dimethyl sulfoxide is frequently associated with toxicity. Complications could be prevented by removing dimethyl sulfoxide from the grafts by washing these before infusion to the patient. Nowadays, automatic washing devices are available offering standardized washing of grafts in closed systems. Compared to manual techniques, the main advantages of these devices are: better efficacy, higher viability of haematopoietic cells, significant reduction of labour, negligible probability of microbial contamination. In 2014, one of these devices, Sepax 2, was purchased by our department.

Patients and methods: Grafts from 5 patients (males, 42–67 years) treated for multiple myeloma (n = 4) and Hodgkin’s lymphoma (n = 1) were thawed and washed before reinfusion using the Sepax 2 (Biosafe S.A. Eysins, Switzerland) automated washing device. Quality controls were performed before cryopreservation and after washing: blood count, cell vitality, number of CD34+ cells and number of CFU-GM (colony-forming units – granulocyte/macrophage).

Results: Concentration of dimethyl sulfoxide before washing of the grafts was 10%, and 0.65% after (0.35–0.69%); recovery of CD34+ cells after washing was 94% (34.57–105.30%); recovery of CFU-GM was 56% (45.14–99.90%); vitality was 85% (78–89%) – the results are listed as median and range. The graft volumes were decreased after washing by 31% (28–42%) compared to the original volumes. Administration of washed grafts was very simple. Patients reported only insignificant inconvenience associated with the transplantations and no significant adverse events were observed. No delay in engraftment was observed: leukocytes > 1 x 109/l day +11, granulocytes > 0,5 x 109/l day +11, PLT > 20 x 109/l day +13 (medians).

Conclusions: The washing of thawed cryopreserved autologous grafts provided required graft volume reduction. The decrease of CD34+ and CFU-GM cells during washing was not significant, cell vitality was satisfactory and the reinfusion of washed grafts was not associated with significant subjective or objective complaints. We conclude that the washing-out of DMSO from grafts using an automated device is feasible and beneficial for both the patients and the staff. The automated washing of dimethyl sulfoxide-containing grafts could be recommended, especially in patients who require lower volumes of reinfused cells.


key words

autologous transplant – haematopoietic stem cell – HSC – Sepax 2 – dimethylsulfoxide – DMSO


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Haematology Internal medicine Clinical oncology
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