Evaluation of intracellular and extracellular trehalose as a cryoprotectant of stem cells obtained from umbilical cord blood.
Cord blood is a source of hematopoietic stem cells used in transplantation in which hematopoietic reconstitution is necessary. This transplant modality requires the cryopreservation of hematopoietic stem cells (HSCs). Dimethyl sulfoxide has been used as a cryoprotectant (CPA) in the cryopreservation of HSCs; however, it has been demonstrated that Me2SO exhibits toxic side effects to the human body. Due to its stability upon freezing, disaccharides such as trehalose have been investigated as a cryoprotectant. This study investigated the hypothesis that a cryopreservation solution containing intracellular and extracellular trehalose improves the recovery of stem cells after cryopreservation. After thawing, the cells were tested for their viability using the 7AAD stain, CD45+/CD34+ cells were assessed using flow cytometry and the MTT viability assay, and the proportion of hematopoietic progenitor cells was measured using the CFU assay. Our results showed the effectiveness of the solution containing intracellular and extracellular trehalose in the cryopreservation of cord blood cells, demonstrating that trehalose may be an optimal cryoprotectant when present both inside and outside of cells.
Histocompatibility and Cryopreservation Laboratory, Department of Histology and Embryology, Institute of Biology Roberto Alcantara Gomes, Rio de Janeiro State University, Pav. Jose Roberto Feresin Moraes, Av Marechal Rondon 381, São Francisco Xavier, 20950-003 Rio de Janeiro, Brazil.
Juliana Pessanha Rodrigues
JP
Motta
Umbilical Cord Blood Bank, Bone Marrow Transplantation Unit, National Cancer Institute, Rio de Janeiro, Brazil.
Flávio Henrique
FH
Paraguassú-Braga
Umbilical Cord Blood Bank, Bone Marrow Transplantation Unit, National Cancer Institute, Rio de Janeiro, Brazil.
Luis Fernando
LF
Bouzas
Histocompatibility and Cryopreservation Laboratory, Department of Histology and Embryology, Institute of Biology Roberto Alcantara Gomes, Rio de Janeiro State University, Pav. Jose Roberto Feresin Moraes, Av Marechal Rondon 381, São Francisco Xavier, 20950-003 Rio de Janeiro, Brazil. Electronic address: lcporto@uerj.br.
Luís Cristóvão
LC
Porto
D002470:Cell Survival; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods; D003451:Cryoprotective Agents / Q000008:administration & dosage / Q000032:analysis / Q000378:metabolism*; D005260:Female; D005312:Fetal Blood / Q000166:cytology*; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D008081:Liposomes; D014199:Trehalose / Q000008:administration & dosage / Q000032:analysis / Q000378:metabolism*
D003451:Cryoprotective Agents; D008081:Liposomes; D014199:Trehalose
Cord blood; Cryopreservation; Liposome; Me(2)SO; Trehalose