Abstract

Histopathologic evaluation and/or archiving of sections of spleen or thymus from all study animals may be mandated by study protocol (e.g., Toxic Substances Control Act-compliant studies). In such cases, whole spleen or thymus is not available for immunophenotyping. It has not been previously demonstrated that immunologic data representative of whole organs can be reliably obtained using a section of the spleen or using one thymic lobe. Light-scatter characteristics and immune cell-surface antigen expression were therefore compared in the right and left halves of the spleen and in the right and left thymic lobes of young adult female C57B1/6 mice and Sprague-Dawley rats. Antigens compared were: mouse spleen – CD11b, CD45R, CD90; rat spleen – CD11b, CD45RA, Pan-T/Ox-52; mouse and rat thymus – CD4, CD8a. There were no significant differences in distribution of cells by size or by expression level for any of these antigens when the right part of the organs was compared to the left part. These data indicate that use of entire spleen or both thymic lobes is not required to reliably quantify resident immune cell subpopulations.

Author Keywords: Spleen; Thymus; Sections; Surface antigens; Mouse; Rat

Article Outline

1. Introduction

2. Methods

2.1. Mice and rats

2.2. Collection of spleen and thymus

2.3. Thymic leukocyte dissociation and isolation

2.4. Splenic dissociation and isolation

2.5. Cell enumeration

2.6. Flow cytometric evaluation of cell-surface markers

2.7. Statistical analysis

3. Results

3.1. Light-scatter analysis of splenocytes and thymocytes

3.2. Splenic cell-surface markers

3.3. Thymic cell-surface markers

4. Discussion

Acknowledgements

References