학술논문
Hydroxyethyl starch as a substitute for dextran 40 for thawing peripheral blood progenitor cell products.
Document Type
Academic Journal
Author
Zhu F; Department of Medicine, Hematology & Oncology Division, Medical College of Wisconsin, Milwaukee, Wisconsin, USA.; Heditke S; Froedtert Hospital, Milwaukee, Wisconsin, USA.; Kurtzberg J; Duke University Medical Center, Durham, North Carolina, USA.; Waters-Pick B; Duke University Medical Center, Durham, North Carolina, USA.; Hari P; Department of Medicine, Hematology & Oncology Division, Medical College of Wisconsin, Milwaukee, Wisconsin, USA.; Margolis DA; Department of Pediatrics, Hematology & Oncology Division, Medical College of Wisconsin, Milwaukee, WI, USA.; Keever-Taylor CA; Department of Medicine, Hematology & Oncology Division, Medical College of Wisconsin, Milwaukee, Wisconsin, USA. Electronic address: ckeever@mcw.edu.
Source
Publisher: Elsevier Country of Publication: England NLM ID: 100895309 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1477-2566 (Electronic) Linking ISSN: 14653249 NLM ISO Abbreviation: Cytotherapy Subsets: MEDLINE
Subject
Language
English
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.
(Copyright © 2015 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.)
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.
(Copyright © 2015 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.)