Related Experiment Videos
Unfractionated human marrow cell cryopreservation using dimethylsulfoxide and hydroxyethyl starch
Cryobiology
|February 1, 1983
Summary
A new cryopreservation method using dimethyl sulfoxide (DMSO) and hydroxyethyl starch (HES) improves bone marrow (BM) cell viability and reduces clumping. This simplified technique enhances cell recovery without requiring specialized freezing equipment or liquid nitrogen storage.
Area of Science:
- Hematology
- Cryobiology
- Cell Biology
Background:
- Cryopreservation of bone marrow (BM) cells is crucial for transplantation and research.
- Conventional methods using 10% dimethyl sulfoxide (DMSO) can lead to cell clumping and reduced viability upon thawing.
- Improving cryopreservation techniques is essential for maximizing functional cell recovery.
Purpose of the Study:
- To evaluate a novel cryopreservation mixture of 5% DMSO and 6% hydroxyethyl starch (HES) for unfractionated BM cells.
- To assess the impact of this DMSO/HES mixture on viable cell yield and post-thaw clumping compared to 10% DMSO alone.
- To determine the efficacy of this simplified cryopreservation method under different storage conditions.
Main Methods:
- Unfractionated BM cells were cryopreserved in 1- to 2-ml aliquots using either 5% DMSO/6% HES or 10% DMSO.
- Cells were stored in the vapor phase of liquid nitrogen for 6 months or at -80°C for 12-16 months.
- Post-thaw analysis included Trypan Blue exclusion for viability and CFU-C (Colony-Forming Unit-Culture) recovery assays.
- Freezing was performed without a rate-controlled apparatus, using sequential -80°C and liquid nitrogen freezers.
Main Results:
- The DMSO/HES mixture significantly increased the percentage of cells excluding Trypan Blue (50% ± 12% vs. 28% ± 12%, P < .01) compared to 10% DMSO.
- CFU-C recovery was higher with the DMSO/HES mixture (110% ± 20% vs. 89% ± 35%, P < .02) compared to 10% DMSO.
- No macroscopic clumping was observed in cells cryopreserved with DMSO/HES, unlike those with 10% DMSO alone.
- BM cells stored at -80°C in DMSO/HES for 12-16 months showed excellent CFU-C recovery (105% ± 39% of prefreeze).
Conclusions:
- The 5% DMSO/6% HES mixture offers a simplified and effective method for cryopreserving unfractionated BM cells.
- This technique enhances viable cell yield and eliminates post-thaw clumping.
- The DMSO/HES method obviates the need for rate-controlled freezers and liquid nitrogen storage for up to a year, making BM cryopreservation more accessible.