Evaluation of trehalose and sucrose as cryoprotectants for hematopoietic stem cells of umbilical cord blood.
Bone marrow transplantation (BMT) is a therapeutic procedure that involves transplantation of hematopoietic stem cells (HSC). To date, there are three sources of HSC for clinical use: bone marrow; mobilized peripheral blood; and umbilical cord blood (UCB). Depending on the stem cell source or type of transplantation, these cells are cryopreserved. The most widely used cryoprotectant is dimethylsulfoxide (Me(2)SO) 10% (v/v), but infusion of Me(2)SO-cryopreserved cells is frequently associated with serious side effects in patients. In this study, we assessed the use of trehalose and sucrose for cryopreservation of UCB cells in combination with reduced amounts of Me(2)SO. The post-thawed cells were counted and tested for viability with Trypan blue, the proportion of HSC was determined by flow cytometry, and the proportion of hematopoeitic progenitor cells was measured by a colony-forming unit (CFU) assay. A solution of 30mmol/L trehalose with 2.5% Me(2)SO (v/v) or 60mmol/L sucrose with 5% Me(2)SO (v/v) produced results similar to those for 10% (v/v) Me(2)SO in terms of the clonogenic potential of progenitor cells, cell viability, and numbers of CD45(+)/34(+) cells in post-thawed cord blood cryopreserved for a minimum of 2 weeks. Thus, cord blood, as other HSC, can be cryopreserved with 1/4 the standard Me(2)SO concentration with the addition of disaccharides. The use of Me(2)SO at low concentrations in the cryopreservation solution may improve the safety of hematopoietic cell transplantation by reducing the side effects on the patient.
Rodrigues
J P
JP
Histocompatibility and Cryopreservation Laboratory, Department of Histology and Embryology, Institute of Biology Roberto Alcantara Gomes, Rio de Janeiro State University, São Francisco Xavier, Brazil.
Paraguassú-Braga
F H
FH
Carvalho
L
L
Abdelhay
E
E
Bouzas
L F
LF
Porto
L C
LC
D002460:Cell Line; D002470:Cell Survival; D003114:Colony-Forming Units Assay; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D005260:Female; D005312:Fetal Blood / Q000166:cytology*; D006412:Hematopoietic Stem Cells / Q000187:drug effects*; D006801:Humans; D020014:K562 Cells; D011247:Pregnancy; D013395:Sucrose / Q000494:pharmacology*; D014199:Trehalose / Q000494:pharmacology*
D003451:Cryoprotective Agents; D013395:Sucrose; D014199:Trehalose; D004121:Dimethyl Sulfoxide