Preclinical evaluation of an automated closed fluid management device: CytomateTM, for washing out DMSO from hematopoietic stem cell grafts after thawing

Abstract

Summary:Infusion of dimethylsulfoxide (DMSO) contained in cryopreserved and thawed hematopoietic stem cell (HSC) grafts is frequently associated with mild or moderate adverse reactions, and occasionally with more severe events including neurological symptoms. The severity of these complications is related to the amount of residual DMSO. We evaluated a recently available, closed, automated and 'cgmp (current good manufacturing practice) compliant' device (CytoMate TM ) for its ability to wash out DMSO at the expense of a limited loss of viable CD34 + cells. A total of 16 procedures were carried out with 39 blood HSC bags intended for destruction. Mean amounts of DMSO for each cellular product (one, two or three bags) were between 12.2 and 39.6 g before thawing; after the washing procedure, residual DMSO quantities were between 0.1 and 3.7 g. When set up to reproducibly allow for a more than 96% elimination of DMSO, processing of thawed cells with the CytoMate TM cell processor resulted in a mean recovery of viable total cells, CD34 + cells and lymphocyte subsets above 60%. We conclude that this simple and efficient washing technique is suitable for routine processing of HSC grafts. Clinical studies will demonstrate whether a reduction in the incidence of adverse effects associated with DMSO infusion is observed.

The Role of High-Dose Chemotherapy Supported by Hematopoietic Stem Cell Transplantation in Patients With Multiple Myeloma: Implications for Nursing

“…Reported adverse reactions to DMSO include nausea and vomiting, abdominal cramps, chills, dyspnea, hypotension or hypertension, bradycardia, arrythmias, alveolar hemorrhage, acute renal failure, and cardiac or respiratory arrest (Alessandrino et al, 1999;Calmels et al, 2003;Hoyt, Szer, & Grigg, 2000;Windrum et al). Aside from toxic reactions to DMSO, acute hemolytic reactions caused by donor-recipient major ABO incompatibility and febrile non-hemolytic reactions may occur.…”

“…The concentrated stem cells then are diluted with an equal volume of human serum albumin solution supplemented with a cryopreservative. Cryopreservation techniques allow for optimal preservation of hematopoietic function and viability of cells (Calmels et al, 2003) Prior to storage, 10% dimethyl sulfoxide (DMSO) is added to the stem cells as a cryoprotectant. DMSO stabilizes cell membranes under rapidly changing conditions and prevents intracellular ice crystal formation during freezing and heat release throughout the period of phase transition, preserving stem cell integrity (Calmels et al;Windrum, Morris, Drake, Niederwieser, & Ruutu, 2005).…”

Autologous Stem Cell Transplantation in the Treatment of Amyloidosis – Can Manipulation of the Autograft Reduce Treatment-Related Toxicity?

“…Lemarie and colleagues used comprehensive automated washing to reach more than 20-fold reduction of DMSO-levels, whereas Syme and coworkers investigated patients with solid tumors and used a relatively extensive manual DMSO depletion (Lemarie et al, 2005;Syme et al, 2004). In the preclinical studies mentioned above (Calmels et al, 2003;Rodriguez et al, 2005), the researhers used a device; CytoMate TM , to perform the washing procedure with phosphate buffered saline (PBS) plus 5% Dextran-40, 5% ACD-A and 1-5% human serum albumin (HSA). While Calmels et al achieved more than 96% elimination of DMSO and a mean recovery of viable total cells, CD34+ cells and lymphocyte subsets above 60%, Rodriguez et al reported 98% DMSO elimination and 103% CD34+ cell recovery.…”

“…Some of the studies where DMSO depletion was utilized, included a large group of patients without bone marrow disease and may not be representative for the present use of autologous peripheral blood stem cell transplantation for hematological malignancies with diffuse infiltration of malignant cells throughout the bone marrow (Calmels et al, 2003;Rodriguez et al, 2005). These preclinical studies investigated grafts of highly selected patients who relapsed or died before autotransplantation could be done (Calmels et al, 2003;Rodriguez et al, 2005). Furthermore, Foïs et al cryopreserved autografts with voluven + DMSO 10% and not DMSO alone, and they did not include detailed analyses of engraftment and transfusion.…”

Assessment of Cryoprotectant Concentration by Electrical Conductivity Measurement and Its Applications in Cryopreservation

“…Therefore, fast and accurate assessment of the CPA concentration in the cell suspension during addition and removal is very important. In order to assess the CPA concentration in cell suspensions, a few methods have been applied by researchers, such as capillary zone electrophoresis [7]), high-performance liquid chromatography (HPLC) [8], and gas chromatography [9]. However, all these approaches are very complex, time-consuming, and expensive.…”