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Concentration of bone marrow progenitor cells by separation on a Percoll gradient using the Haemonetics model 30
Y Humblet1, P Lefebvre, J L Jacques
1Ludwig Institute for Cancer Research, Brussels, Belgium.
Bone Marrow Transplantation
|January 1, 1988
Summary
This study introduces a two-step bone marrow processing method using a Haemonetics device and Percoll density gradient, significantly reducing red blood cells and improving progenitor cell recovery for effective ex vivo purging and transplantation.
Area of Science:
- Hematology
- Cellular Biology
- Biotechnology
Background:
- Optimal ex vivo bone marrow purging requires concentrating progenitor cells while removing red blood cells and polymorphonuclear leukocytes.
- The Haemonetics model 30 alone is insufficient for achieving optimal bone marrow processing goals.
Purpose of the Study:
- To develop and evaluate a two-step procedure combining the Haemonetics model 30 with a density gradient for enhanced bone marrow processing.
- To improve the efficiency of ex vivo bone marrow purging for transplantation.
Main Methods:
- A two-step procedure was implemented: first, obtaining the buffy coat from bone marrow grafts; second, reintroducing buffy coats into the Haemonetics bowl with Percoll (1.079 g/ml) for mononuclear cell recovery.
- The method focused on recovering light density mononuclear cells.
Main Results:
- The two-step procedure significantly reduced marrow volume (to 9%), red blood cell contamination (to 0.96%), and nucleated cell contamination (to 16%) compared to single-step methods.
- Colony-forming unit-granulocyte-macrophage (CFU-GM) recovery remained high at 70% after the second step.
- Of 24 patients grafted, 13 showed complete and rapid hematological recovery.
Conclusions:
- Combining a density gradient with the Haemonetics model 30 offers an effective method for ex vivo bone marrow purging.
- This optimized procedure reduces cellular contaminants and volume, facilitating purging without compromising graft capability.