Post-thaw dimethyl sulfoxide reduction in autologous peripheral blood progenitor cell suspensions

Jandová, Miroslava; Měřička, Pavel; Gregor, Jiří; Lánská, Miriam; Bezrouk, Aleš; Čížková, Dana; Radocha, Jakub

doi:10.1016/j.htct.2025.103965

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Original article • Hematol., Transfus. Cell Ther. 47 (4) • 2025 • https://doi.org/10.1016/j.htct.2025.103965 linkcopy

Post-thaw dimethyl sulfoxide reduction in autologous peripheral blood progenitor cell suspensions

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

Background and objectives Dimethyl sulfoxide has become the most common cryoprotectant used for cryopreservation of hematopoietic progenitor cells because of its efficiency, regardless of its potentially toxic side effects. Its application is considered safe, provided that the daily dose administered does not exceed 1 gram per kilogram of patient weight. Indications for its reduction after thawing are limited to patients with high risk of malignant arrhythmia and those with severely impaired renal function. However, dimethyl sulfoxide reduction can lead to the loss of viable progenitors.

Methods A retrospective study of viable hematopoietic progenitor cell recovery after dimethyl sulfoxide reduction was performed with 13 patients (nine men, four women) with secondary amyloidosis in multiple myeloma (n = 9), primary amyloid light chain amyloidosis (n = 3), or severe adverse reaction at the beginning of the hematopoietic progenitor cell concentrate infusion (n = 1). The Wilcoxon signed-rank test was used.

Results The results of the dimethyl sulfoxide reduction process showed a high recovery of viable nucleated cells (median: 120.85 %), and of viable mononuclear cells (median: 104.53 %). There was a significant decrease in total number of viable CD34⁺ cells in comparison with data obtained after original collection (median: 51.49 %). No significant decrease in colony-forming unit capacity was observed after dimethyl sulfoxide reduction (median: 93.37 %).

Conclusion The dimethyl sulfoxide removal process and total process recoveries revealed considerable individual variability. To minimize the risk of prolonged engraftment or non-engraftment, it is important to apply this process only to high-risk patients.

Keywords
Peripheral blood progenitor cells; Cryopreservation; DMSO reduction; Amyloidosis; Cell recovery

Patient number	Sex	Age (year)	Weight (kg)	Diagnosis/reason for washing
1	male	55	81	Multiple myeloma - secondary amyloidosis - suspected amyloidosis of heart
2	female	61	52	Primary amyloidosis of bone marrow, liver, and kidneys
3	male	48	94	Multiple myeloma - allergic adverse reaction (DMSO)
4	male	56	91	Multiple myeloma - secondary amyloidosis - suspected amyloidosis of heart
5	female	55	80	Multiple myeloma - secondary amyloidosis of kidneys
6	female	57	61	Multiple myeloma - secondary amyloidosis - suspected amyloidosis of heart
7	male	69	93	Primary amyloidosis of liver and lungs
8	female	51	58	Multiple myeloma - secondary amyloidosis of gastrointestinal tract and kidneys
9	male	44	102	Multiple myeloma - secondary amyloidosis - suspected amyloidosis of heart
10	male	58	84	Multiple myeloma - secondary amyloidosis of duodenum and heart
11	male	66	77	Multiple myeloma - secondary amyloidosis of gastrointestinal tract
12	male	59	67	Primary amyloidosis of bone marrow and kidneys
13	male	70	103	Multiple myeloma - secondary amyloidosis - suspected amyloidosis of heart

Parameter	Pre-process (leukapheresis) median (Q1; Q3)	Post-process (freezing-thawing) median (Q1; Q3)	p-value
TNC/kg (x 10⁸)	2.46 (1.96; 4.01)	1.79 (1.24; 3.04)	0.006*
NC viability ( %)	100 (100; 100)	82 (74; 95)	0.002*
TMNC/kg (x 10⁸)	1.51 (0.80; 1.78)	1.40 (1.01; 2.09)	0.364
MNC viability ( %)	100 (100; 100)	87 (92; 99)	0.002*
MNC from TNC ( %)	56 (33; 66)	70 (60; 83)	0.002*
CD34⁺/kg (x 10⁶)	3.71 (1.64; 6.15)	3.88 (1.42; 6.84)	0.529
CD34⁺ from TNC ( %)	1.33 (0.80; 2.00)	1.63 (0.87; 2.09)	0.014*
CFU-GM/kg (x 10⁵)	2.97 (1.71; 4.05)	1.68 (1.42; 2.91)	0.002*

Parameter	Pre-process (freezing-thawing) median (Q1; Q3)	Post-process (DMSO removal) median (Q1; Q3)	P value
TNC/kg (x 10⁸)	1.79 (1.24; 3.04)	1.79 (1.37; 2.20)	0.576
NC viability ( %)	82 (74; 95)	78 (69; 85)	0.025
TMNC/kg (x 10⁸)	1.40 (1.01; 2.09)	1.40 (0.87; 1.75)	0.402
MNC viability ( %)	87 (92; 99)	87 (83; 92)	0.002*
MNC from TNC ( %)	70 (60; 83)	63 (47; 83)	0.081
CD34⁺/kg (x 10⁶)	3.88 (1.42; 6.84)	1.76 (1.13; 3.65)	0.018
CD34⁺ from TNC ( %)	1.63 (0.87; 2.09)	0.90 0.51; 1.51)	0.002*
CFU-GM/kg (x 10⁵)	1.68 (1.42; 2.91)	1.82 (1.31; 2.71)	0.133

Parameter	Pre-process (leukapheresis) median (Q1; Q3)	Post-process (DMSO removal) median (Q1; Q3)	P value
TNC/kg (10⁸)	2.46 (1.96; 4.01)	1.79 (1.37; 2.20)	0.036
NC viability (%)	100 (100; 100)	78 (69; 85)	0.002*
TMNC/kg (10⁸)	1.51 (0.80; 1.78)	1.40 (0.87; 1.75)	0.944
MNC viability (%)	100 (100; 100)	87 (83; 92)	0.002*
MNC from TNC (%)	56 (33; 66)	63 (47; 83)	0.036
CD34⁺/kg (10⁶)	3.71 (1.64; 6.15)	1.76 (1.13; 3.65)	0.010*
CD34⁺ from TNC (%)	1.33 (0.80; 2.00)	0.90 0.51; 1.51)	0.003*
CFU-GM/kg (10⁵)	2.97 (1.71; 4.05)	1.82 (1.31; 2.71)	0.006*

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Associação Brasileira de Hematologia, Hemoterapia e Terapia Celular (ABHH) R. Dr. Diogo de Faria, 775 cj 133, 04037-002, São Paulo / SP - Brasil - São Paulo - SP - Brazil
E-mail: htct@abhh.org.br

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[TFN1] ^⁎
Statistically significant difference

TNC: total number of viable nucleated cells; NC: nucleated cells; TMNC: total number of viable mononuclear cells; MNC mononuclear cells; CFU-GM: colony forming unit-granulocyte macrophage.