Hydroxyethyl starch as a substitute for dextran 40 for thawing peripheral blood progenitor cell products

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• 作者：Fenlu Zhu¹ ; Sarah Heditke² ; Joanne Kurtzberg³ ; Barbara Waters-Pick³ ; Parameswaran Hari¹ ; David A. Margolis⁴ ; Carolyn A. Keever-Taylor¹ ; ^{ckeever@mcw.edu" class="auth_mail" title="E-mail the corresponding author}
• 关键词：cryopreservation ; thawing ; low molecular weight dextran ; dextran 40 ; hydroxyethyl starch
• 刊名：Cytotherapy
• 出版年：2015
• 出版时间：December 2015
• 年：2015
• 卷：17
• 期：12
• 页码：1813-1819
• 全文大小：531 K

文摘

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.