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Isolation of Myeloid Dendritic Cells and Epithelial Cells from Human Thymus
Published on: September 20, 2013
CD34+CD38-lin- cord blood cells develop into dendritic cells in human thymic stromal monolayers and thymic nodules
G D Miralles1, C A Smith, L P Whichard
1Department of Medicine, Duke University Medical Center, Durham, NC 27710, USA.
Insights
Human thymic stroma supports the development of primitive hematopoietic stem cells into mature dendritic cells (DCs). This study provides a novel system for understanding DC development in the thymus.
Area of Science:
- Immunology
- Cell Biology
- Developmental Biology
Background:
- Thymic dendritic cells (DCs) possess unique properties distinct from DCs in other tissues.
- Understanding human thymic DCs is limited due to challenges in their isolation and in vitro culture.
Purpose of the Study:
- To investigate the capacity of human thymic stroma to support the development of hematopoietic stem cells into mature DCs.
- To establish novel in vitro culture systems for studying human thymic DC ontogeny.
Main Methods:
- Coculture of CD34+CD38-lin- and CD34+CD38+lin- umbilical cord blood cells with human thymic stromal monolayers.
- Utilizing a novel tissue culture system with thymic nodules comprising epithelial cells and fibroblasts.
Main Results:
- Significant expansion (43-fold and 32-fold) of umbilical cord blood progenitors was observed.
- Generated cells exhibited mature DC morphology, phenotype (MHC class I/II, CD1a, CD11c, CD80, CD86, etc.), and function.
- Demonstrated potent allogeneic T cell activation by the generated DCs.
Conclusions:
- Human thymic stroma efficiently supports the differentiation of primitive hematopoietic stem cells into mature DCs.
- The described culture systems provide valuable tools for studying human DC development within thymic microenvironments.
Abstract:
Thymic dendritic cells (DCs) appear to have distinct biologic and functional properties compared with DCs in other tissues. Currently, little is known about human thymic DCs because they have been difficult to isolate and culture in vitro. Here, we report that human thymic stroma can support the development of primitive human hemopoietic stem cells into mature DCs without cytokine or serum supplementation. Coculture of CD34+CD38-lineage (lin)- and CD34+CD38+lin- umbilical cord blood cells with thymic stromal monolayers induced 43 +/- 17-fold and 32 +/- 16-fold expansions, respectively, of umbilical cord blood progenitors and also generated large numbers of cells with the morphologic, phenotypic, and functional characteristics of mature DCs. These cells expressed class I and class II MHC, CD1a, CD2, CD4, CD11c, CD40, CD45, CD80, CD83, and CD86 and were potent stimulators of allogeneic T cell activation. Primitive hemopoietic progenitors also developed into mature DCs in a novel tissue culture system of thymic nodules wherein thymic epithelial cells and fibroblasts were grown in nodular aggregates in vitro. These results demonstrate that human thymic stroma efficiently supports the development of CD34+CD38-lin- cord blood cells into mature DCs. In addition, the culture conditions described in this report are useful systems for studying the ontogeny of human DCs in thymic microenvironments.
