Characterization of Thymic Nurse-Cell Lymphocytes, Using an Improved Procedure for Nurse-Cell Isolation

Mireille Lahoud, David Vremec, Richard L. Boyd, Ken Shortman

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Thymic nurse cells (TNC), multicellular complexes consisting of lymphoid cells enclosed within cortical epithelial cells, were isolated from mouse thymus by a modified procedure allowing immunofluorescent labeling and flow cytometric analysis of their lymphoid contents (TNC-L). Collagenase was the only protease used for tissue digestion, to ensure that surface antigen markers remained intact. Zonal unit-gravity elutriation was used to enrich the TNC on the basis of their high sedimentation rate, followed by immunomagnetic bead depletion to remove residual mononuclear cell contaminants and a density separation to remove debris. The TNC-L were then released from inside TNC by a short period of culture. The measured contamination of TNC-L with exogenous thymocytes was around 0.5%. Three-color immunofluorescent labeling revealed that TNC-L included, as well as a majority of immature CD4+8+3low thymocytes, about 12% of apparently mature CD4+83high and CD4~8+3high thymocytes. TNC are located in the cortex, where mature cells are rare; the occurrence of mature phenotype cells within these structures suggests that they represent a microenvironment for the selection and generation of mature T cells.

Original languageEnglish
Pages (from-to)103-112
Number of pages10
JournalDevelopmental Immunology
Issue number2
Publication statusPublished - 1993


  • cell elutriation
  • cell separation
  • multicellular complexes
  • positive selection
  • T-cell maturation
  • Thymic nurse cells
  • thymus microenvironments

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