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Updated: Jun 6, 2026
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Cryopreservation and Bioenergetic Evaluation of Human Peripheral Blood Mononuclear Cells
Published on: October 20, 2023
[Comparison of different cryopreservation systems for peripheral blood stem cells]
You-Zhang Huang1, Jian-Liang Shen, Ping-Di Yang
1Department of Hematology, Navy General Hospital of PLA, Beijing 100037, China.
Summary
The combination of 5% dimethyl sulfoxide (DMSO) and 6% hydroxyethyl starch (HES) is a superior cryopreservative for hematopoietic stem cells compared to 10% DMSO. For cryopreservation under one year, a -80°C refrigerator is effective, while liquid nitrogen is recommended for longer durations.
Area of Science:
- Cellular Biology
- Cryobiology
- Hematology
Background:
- Hematopoietic stem cell (HSC) transplantation is a critical therapy.
- Effective cryopreservation is essential for HSC viability and function.
- Optimizing cryopreservative systems is crucial for successful HSC banking and clinical application.
Purpose of the Study:
- To evaluate different cryopreservative systems for hematopoietic stem cells.
- To compare the efficacy of dimethyl sulfoxide (DMSO) alone versus a DMSO and hydroxyethyl starch (HES) combination.
- To assess the impact of different cooling and storage methods on HSC recovery.
Main Methods:
- Peripheral blood mononuclear cells from 20 individuals were used.
- Cells were treated with either 10% DMSO or a combination of 5% DMSO and 6% HES.
- Cooling was performed using a -80°C refrigerator (Refr) or a programmed cryogenic system (PCS).
- Storage was in Refr or liquid nitrogen, with assays for GM-CFU, LTC-IC, CD34+ cells, and trypan blue resistance (TBR).
Main Results:
- The 5% DMSO-6% HES combination yielded significantly higher recovery rates for CFU-GM, LTC-IC, CD34+ cells, and TBR compared to 10% DMSO (P < 0.05).
- No significant difference in recovery rates was observed between Refr and Refr with PCS cooling methods.
- Cryopreservation for less than one year showed no significant difference in recovery rates across Refr, Refr with PCS, and liquid nitrogen storage.
- Recovery rates decreased significantly after two years of cryopreservation in the Refr.
Conclusions:
- 5% DMSO-6% HES is a more effective cryopreservative for HSCs than 10% DMSO.
- Refr cryopreservation is suitable for HSCs stored for up to one year.
- Liquid nitrogen cryopreservation is recommended for storage exceeding one year.
- Immediate dilution or removal of cryopreservatives post-thawing is vital to maintain cell activity.
