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[A Simplified Method for Cryopreservation of Peripheral Blood Stem Cells]
Miao Zhang1, Bo-Long Zhang, Zuo-Yun Zhang
1Department of Hematology and Institute of Clinical Basic Medical Sciences, The General Hospital of PLA, Beijing 100853, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi
|February 13, 2003
Summary
A simplified cryopreservation method for peripheral blood stem cells (PBSCs) using 5% DMSO, 3% HES, and 4% HSA at -80°C offers higher recovery rates and reduced toxicity compared to traditional methods. This technique is ideal for facilities lacking specialized freezing equipment.
Area of Science:
- Hematology
- Cell Biology
- Cryobiology
Background:
- Traditional cryopreservation of peripheral blood stem cells (PBSCs) at -196°C requires rate-controlled freezing and 10% dimethyl sulfoxide (DMSO).
- This method can be complex and inaccessible for institutions lacking specialized equipment.
- High DMSO concentrations can lead to toxicity in patients.
Purpose of the Study:
- To develop and evaluate a simplified cryopreservation technique for PBSCs.
- To compare the efficacy of the simplified method with traditional cryopreservation.
- To assess the impact of reduced DMSO concentration and storage temperature on cell viability and recovery.
Main Methods:
- Cryopreservation of PBSCs using a simplified method with 5% DMSO, 3% hydroxyethyl starch (HES), and 4% human serum albumin (HSA) at -80°C.
- Comparison of cell viability and recovery rates of mononuclear cells (MNC), CFU-GM, and CD34(+) cells with traditional methods.
- Assessment of long-term storage (up to 24 months) and thawing parameters (37°C vs. 20°C).
Main Results:
- The simplified method yielded higher MNC and CFU-GM recovery rates without agglutination.
- Satisfactory recovery rates for CFU-GM and CD34(+) cells were achieved after 24 months of storage at -80°C.
- No significant difference in cryopreserved PBSC parameters was observed between thawing at 37°C and 20°C.
Conclusions:
- The simplified cryopreservation technique is simpler and more effective than the traditional method.
- This method is suitable for institutions without rate-controlled freezing facilities.
- Reduced DMSO concentration decreases toxicity, making it safer for patients.