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Morphologic and functional characteristics of alveolar macrophages following cryopreservation
L S Lamb1, J B Willoughby, W F Willoughby
1Department of Pathology, School of Medicine, University of South Carolina, Columbia 29203, USA.
Abstract:
Cryopreservation of nucleated blood and bone marrow mononuclear cells has been used both to preserve such cells for clinical, diagnostic, or research use and to eliminate them as passengers in frozen tissue destined for transplantation. Although techniques for macrophage and monocyte cryopreservation have been described, little work has been done on the functional state of these cells following frozen storage. Using rabbit alveolar macrophages (AM), we have shown that AM can be frozen, stored, and recovered without morphologic changes. Furthermore, freshly isolated and cryopreserved AM do not differ in their adherence characteristics or the organization of their actin cytoskeleton. Cryopreserved normal AM also retained inducible functions such as superoxide anion (O2-) release and production of tumor necrosis factor (TNF) and interleukin-1 (IL-1). In order to further define the effects of cryopreservation on macrophage biology, we have also frozen macrophages which have been primed in vitro, and have demonstrated that the primed state remains at prefreezing levels after thawing and subsequent analysis. These data help define the effects of cryopreservation on cell function and establish parameters for the clinical and experimental evaluation of cryopreserved mononuclear phagocytes.
Insights
Cryopreservation preserves rabbit alveolar macrophages (AM) without altering morphology or function. Cryopreserved AM retain adherence, cytoskeleton organization, and inducible functions like cytokine release, proving their viability for research and clinical use.
Area of Science:
- Immunology
- Cell Biology
- Cryobiology
Background:
- Cryopreservation is vital for preserving mononuclear cells for clinical and research applications.
- Limited data exists on the functional integrity of cryopreserved macrophages post-thaw.
Purpose of the Study:
- To evaluate the effects of cryopreservation on the morphology and function of rabbit alveolar macrophages (AM).
- To determine if cryopreservation impacts macrophage adherence, cytoskeleton, and inducible functions.
Main Methods:
- Rabbit alveolar macrophages (AM) were cryopreserved and thawed.
- Morphological integrity, adherence characteristics, and actin cytoskeleton organization were assessed.
- Inducible functions, including superoxide anion release and cytokine production (TNF, IL-1), were measured.
- Primed macrophages were cryopreserved to assess functional memory retention.
Main Results:
- Cryopreservation and thawing did not induce morphological changes in AM.
- Cryopreserved AM exhibited similar adherence and actin cytoskeleton organization compared to fresh cells.
- Inducible functions, including superoxide anion release and production of tumor necrosis factor (TNF) and interleukin-1 (IL-1), were retained.
- The primed state of macrophages was preserved after cryopreservation and thawing.
Conclusions:
- Cryopreservation is a suitable method for storing rabbit alveolar macrophages (AM) without compromising their key functions.
- Cryopreserved AM maintain their viability, adherence, cytoskeleton, and inducible functions, making them valuable for research and clinical applications.
- These findings establish parameters for evaluating cryopreserved mononuclear phagocytes.