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Density gradient isolation of peripheral blood mononuclear cells using a blood cell processor
1American Red Cross, Biomedical Research and Development Laboratory, Rockville, Maryland.
Transfusion
|March 1, 1988
Summary
The Haemonetics V50 processor efficiently isolates mononuclear cells (MNC) for stem cell therapy, offering higher yields and purity compared to the Cobe 2991. This method preserves hematopoietic progenitor cell function, making it a cost-effective and sterile option.
Area of Science:
- Hematology
- Cellular Biology
- Biomedical Engineering
Background:
- Hematologic reconstitution using peripheral blood stem cells requires large numbers of mononuclear cells (MNC).
- Efficient isolation of MNC is crucial for successful stem cell therapies.
Purpose of the Study:
- To evaluate the efficacy of two blood cell processors, Haemonetics V50 (V50) and Cobe 2991 (2991), for isolating MNC via Ficoll-diatrizoate density gradient centrifugation.
- To compare the yield, purity, and functional integrity of hematopoietic progenitor cells obtained from both systems.
Main Methods:
- Discontinuous Ficoll-diatrizoate density gradient centrifugation was performed using the V50 and 2991 processors.
- Mononuclear cells were isolated from peripheral blood buffy coats.
- Cell recovery and purity (lymphocytes and monocytes among leukocytes) were assessed.
- Cloning efficiencies of hematopoietic progenitor cells (CFU-GM and BFUe) were determined.
Main Results:
- The V50 processor achieved higher MNC yield (92%) compared to the 2991 (75%) for larger cell loads (3-7 X 10(9) MNC).
- Initial separation in the V50 with a smaller cell load (6.23 X 10(8) MNC) resulted in 75% recovery and 95% purity.
- No significant difference in the cloning efficiency of hematopoietic progenitor cells was observed between the V50 and 2991, or with standard gradient centrifugation in tubes.
- The V50 allowed for combined leukapheresis and MNC separation in a single procedure, processing an average of 6 X 10(9) MNC with 82% recovery and 95% purity.
Conclusions:
- The Haemonetics V50 processor is a suitable and potentially superior alternative to the Cobe 2991 for purifying MNC from peripheral blood.
- The V50 offers advantages in terms of operational simplicity, cost-effectiveness, and maintenance of sterility.
- The method preserves the functional capacity of essential hematopoietic progenitor cells for therapeutic applications.