Faculty Collaboration Database

Hydroxyethyl starch as a substitute for dextran 40 for thawing peripheral blood progenitor cell products. Cytotherapy 2015 Dec;17(12):1813-9

Date

10/12/2015

Pubmed ID

26454752

DOI

10.1016/j.jcyt.2015.08.007

Scopus ID

2-s2.0-84947594247 (requires institutional sign-in at Scopus site)   15 Citations

Abstract

BACKGROUND AIMS: Removing DMSO post-thaw results in: reduced infusion reactions, improved recovery and stability of viable CD34+ cells. Validated methods use 5%-8.3% Dextran 40 with 2.5%-4.2% HSA for this purpose. Recent shortages of clinical grade Dextran require identification of suitable alternatives.

METHODS: PBPC were used to compare a standard 2X wash medium of 5 parts 10% Dextran 40 in saline (DEX) with 1 part 25% HSA (8.3% DEX/ 4.2% HSA) with Hydroxyethyl Starch (HES)-based solutions. Cells in replicate bags were diluted with an equal volume of wash solution, equilibrated 5 minutes, the bag filled with wash medium, pelleted and the supernatant expressed. Bags were restored to the frozen volume in wash medium and tested by single platform flow cytometry and CFU. Total viability, viable TNC, MNC, and CD34+ cell recovery, and CD34+ cell viability were compared immediately post-thaw and after 90 minutes.

RESULTS: 5.2% HES/4.2% HSA did not differ from our standard in CD34 recovery or viability. Due to concerns that high concentrations of HES could affect renal function we tested 0.6% HES/2.5% HSA resulting in significantly poorer CD34 recovery and viability. Results improved using 2.4% HES/4.2% HSA and when 0.6% HES/4.2%HSA was used no significant differences were seen. CFU assays confirmed no differences between the standard dextran arm and HES at 2.4% or 0.6% so long as HSA was at 4.2%.

CONCLUSIONS: We conclude that HES from 0.6% to 5.2% with 4.2% HSA is a suitable substitute for Dextran 40 as a reconstitution/washing medium for PBPC products.

Author List

Zhu F, Heditke S, Kurtzberg J, Waters-Pick B, Hari P, Margolis DA, Keever-Taylor CA

Authors

Parameswaran Hari MD Adjunct Professor in the Medicine department at Medical College of Wisconsin
David A. Margolis MD Chair, Professor in the Pediatrics department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Cell Survival
Cells, Cultured
Colony-Forming Units Assay
Cryopreservation
Culture Media
Dextrans
Freezing
Hematopoietic Stem Cells
Humans
Hydroxyethyl Starch Derivatives
Male