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Culture of Macrophage Colony-stimulating Factor Differentiated Human Monocyte-derived Macrophages
Published on: June 30, 2016
Sterile isolation and cryopreservation of human adherent monocytes/macrophages from mononuclear cell apheresis
Stephen J Wagner1, Andrew Myrup, Yanwen Xu
1American Red Cross, Biomedical Services, Jerome H. Holland Laboratory for the Biomedical Sciences, Blood & Cell Therapy Development, Rockville, Maryland 20855, USA. wagners@usa.redcross.org
Abstract:
There is evidence from animal and human study that suggests clinical use of monocytes/macrophages may be of benefit in wound healing, tissue regeneration, and cancer therapy. To facilitate further study, a method was developed for sterile isolation and cryopreservation of adherent monocyte/macrophages from mononuclear cell apheresis units collected from unstimulated normal human donors. Preparations contained approximately 1 x 108 total cells and were comprised of approximately 60% monocytes, 38% lymphocytes, and 2% granulocytes. Cells could be cryopreserved for up to 8 months and subsequently thawed and stored at 1-6 degrees C for up to 4 hours with retention of viability and adequate phagocytic function. These cells can be used in clinical trials to determine their possible therapeutic benefit, e.g., whether administration of exogenously supplied cells improves the healing of chronic wounds or promotes the regeneration of transected nerves.
Insights
A new method allows sterile isolation and cryopreservation of human monocytes/macrophages for therapeutic applications. These cells show viability and function after storage, supporting potential use in wound healing and tissue regeneration clinical trials.
Area of Science:
- Immunology
- Cell Biology
- Regenerative Medicine
Background:
- Monocytes/macrophages show therapeutic potential in wound healing, tissue regeneration, and cancer therapy.
- Clinical application requires reliable methods for cell isolation and preservation.
Purpose of the Study:
- To develop a method for sterile isolation and cryopreservation of human monocytes/macrophages.
- To assess the viability and function of cryopreserved cells for potential clinical use.
Main Methods:
- Isolation of adherent monocytes/macrophages from mononuclear cell apheresis units.
- Sterile processing and cryopreservation of isolated cells.
- Assessment of cell viability and phagocytic function post-thaw and short-term storage.
Main Results:
- Developed a method yielding approximately 1 x 10^8 cells per preparation, with ~60% monocytes.
- Cryopreservation up to 8 months maintained cell viability and phagocytic function.
- Thawed cells retained viability and function for up to 4 hours at 1-6°C.
Conclusions:
- A reproducible method for sterile isolation and cryopreservation of human monocytes/macrophages is established.
- Cryopreserved monocytes/macrophages are suitable for clinical trials investigating therapeutic benefits.
- These cells hold promise for enhancing chronic wound healing and nerve regeneration.
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