5% Dimethyl sulfoxide (DMSO) and pentastarch improves cryopreservation of cord blood cells over 10% DMSO

• Published in: Transfusion (Philadelphia, Pa.) Vol. 50; no. 10; pp. 2158 - 2166
• Main Authors: Hayakawa, Jun, Joyal, Elizabeth G., Gildner, Jean F., Washington, Kareem N., Phang, Oswald A., Uchida, Naoya, Hsieh, Matthew M., Tisdale, John F.
• Format: Journal Article
• Language: English
• Published: Malden, USA Blackwell Publishing Inc 01-10-2010; Wiley
• Subjects: Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Animals; Antigens, CD34 - metabolism; Biological and medical sciences; Blood Preservation - methods; Blood. Blood and plasma substitutes. Blood products. Blood cells. Blood typing. Plasmapheresis. Apheresis; Bone marrow, stem cells transplantation. Graft versus host reaction; Cell Survival - drug effects; Cryopreservation - methods; Cryoprotective Agents - adverse effects; Dimethyl Sulfoxide - adverse effects; Fetal Blood - cytology; Humans; Hydroxyethyl Starch Derivatives - adverse effects; Leukocytes, Mononuclear - cytology; Leukocytes, Mononuclear - transplantation; Male; Medical sciences; Mice; Mice, Inbred NOD; Mice, SCID; Transfusions. Complications. Transfusion reactions. Cell and gene therapy; Non steroidal antiinflammatory agent; Prostaglandin-endoperoxide synthase; Transfusion; Enzyme; Cryopreservation; Dimethyl sulfoxide; Enzyme inhibitor; Oxidoreductases
• ISSN: 0041-1132; 1537-2995
• DOI: 10.1111/j.1537-2995.2010.02684.x

BACKGROUND: Cell number and viability are important in cord blood (CB) transplantation. While 10% dimethyl sulfoxide (DMSO) is the standard medium, adding a starch to freezing medium is increasingly utilized as a cytoprotectant for the thawing process. Similar to hetastarch, pentastarch has the advantages of faster renal clearance and less effect on the coagulation system.
STUDY DESIGN AND METHODS: We compared a lower DMSO concentration (5%) containing pentastarch with 10% DMSO and performed cell viability assay, colony‐forming units (CFUs), and transplantation of CB cells in NOD/SCID IL2Rγnull mice.
RESULTS: CB cells in 5% DMSO/pentastarch had similar CD34+, CD3+, and CD19+ cell percentages after thawing as fresh CB cells. CB cells in 5% DMSO/pentastarch had higher viability (83.3 ± 9.23%) than those frozen in 10% DMSO (75.3 ± 11.0%, p < 0.05). We monitored cell viability postthaw every 30 minutes. The mean loss in the first 30 minutes was less in the 5% DMSO/pentastarch group. At the end of 3 hours, the viability decreased by a mean of 7.75% for the 5% DMSO/pentastarch and 17.5% for the 10% DMSO groups. CFUs were similar between the two cryopreserved groups. Frozen CB cells engrafted equally well in IL2Rγnull mice compared to fresh CB cells up to 24 weeks, and CB cells frozen in 5% DMSO/pentastarch engrafted better than those in 10% DMSO.
CONCLUSION: Our data indicate that the lower DMSO concentration with pentastarch represents an improvement in the CB cryopreservation process and could have wider clinical application as an alternate freezing medium over 10% DMSO.