Abstract
The thymus is a very complex organ that regulates T-cell production Thymocytes (immature T-cells) constitute by far the largest cellular population in the organ (several billions of thymocytes in a child’s thymus), but small numbers of other hematopoietic cells are found in the intrathymic microenvironment, such as thymic monocytes, macrophages, interdigitating dendritic cells, and B-cells Stromal cells of nonhematopoietic origm comprise thymic epithelial cells (TEC) organized in a network throughout the organ, thymic fibioblasts of the capsule and interlobular septae, and endothelial cells of the thymic vasculature The proportion of these thymic cellular components varies with age (1). Fat infiltration becomes significant at puberty and increases throughout adulthood. Intrathymic T-cell maturation is supported in part by TEC, which express cell-surface molecules interacting with counterreceptors on the maturing thymocytes (2–4) Importantly, TEC induce positive selection and major histocompatibility complex restriction of T-cells (5) In vitro studies have shown that TEC produce numerous cytokines (4, 6, 7), which may directly and/or indirectly contribute to T-cell maturation. Reciprocally, TEC functions are affected by interactions with T-cells (t4, 8). Monolayer cultures of TEC provide in vitro systems to study the biology of TEC (2–7) However, investigators should be aware that monolayer cultures may not be representative of a three-dimensionally structured TEC network, and the function of isolated cells may differ from that of cells in the midst of a complex environment One method to obtain highly purified monolayer cultures of human TEC is described here Nontransformed human TEC can be propagated and passaged up to six times before cells become senescent. Careful “budgeting” of the cell stocks by freezing early passages can provide a long-lasting supply of purified TEC strains with determined purity
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Nakahama, M., Mohri, N., Mori, S., Shindo, G., Yokoi, Y., Machinami, R., (1990) Immunohistochemical and histometrical studies of the human thymus with special emphasis on age-related changes in medullary epithelial and dendritic cells Virchows Arch B Cell Pathol 58, 245–251
Singer, K. H., Denning, S. M., Whichard, L. P., Haynes, B. F., (1990) Thymocyte LFA-1 and thymic epithelial cell ICAM-1 molecules mediate binding of activated human thymocytes to thymic epithelial cells J Immunol 144, 2931
Galy, A. H. M., Spits, H., (1992) CD40 is functionally expressed on human thymic epithelial cells J Immunol 149, 775–782
Galy, A. H. M., Spits, H., (1991) IL-l, IL-4 and IFN-Ύ differentially regulate cytokine production and cell surface molecule expression in cultured human thymic epithelial cells J Immunol 147, 3823–3830
Martin Foncheca, A., Schuurman, H. J., and Zapata, A., (1994) Role of thymic stromal cells in thymocyte education. a comparative analysts of different models Thymus 22, 201–213
Galy, A. H. M., de Waal Malefyt, R., Barcena, A., Mohan-Peterson, S., Spits, H., (1993) Untransfected and SV40-transfected fetal and postnatal human thymic stromal cells analysis of phenotype, cytokine gene expression and cytokine production Thymus 22, 13–33
Galy, A. H. M., Spits, H., and Hamilton, J. A., (1993) Regulation of M-CSF production by cultured human thymic epithelial cells. Lymphokine Cytokine Res 12, 265–270
Surh, C. D., Ernst, B., Sprent, J., (1992) Growth of epithelial cells in the thymic medulla is under the control of mature T cells J Exp Med 176, 611–616
Sun, L., Serrero, G., Piltch, A., and Hayashi, J., (1987) EGF receptors on TEA3AI endocrine thymic epithelial cells Biochem Biophys Res Commun 148, 603–608.
Hadden, J. W., Galy, A., Chen, H., Hadden, E. M., (1989) A pituitary factor Induces thymic epithelial cell proliferation in vitro Brain Behav Immun 3, 149–159
Singer, K. H., Harden, E. A., Robertson, A. L., Lobach, D. F., and Haynes, B. F., (1985) In vitro growth and phenotypic characterization of mesodermal-dertved and epnhehal components of normal and abnormal thymus Hum Immunol 13, 16l–176
Moll, R., Franke, W. W., Schiller, D. L., Getger, B., and Krepler R., (1982) The catalog of human cytokeratins patterns of expression in normal epithelia, tumors and cultured cells Cell 31, 11
Anderson, G., Jenkinson, E. J., Moore, N. C., and Owen, J. J. T., (1993) MHC class II positive epithelium and mesenchyme cells are both required for T-cell development in the thymus Nature 362, 70–73
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1996 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Galy, A.H.M. (1996). Human Thymic Epithelial Cell Cultures. In: Jones, G.E. (eds) Human Cell Culture Protocols. Methods in Molecular Medicine, vol 2. Humana Press. https://doi.org/10.1385/0-89603-335-X:111
Download citation
DOI: https://doi.org/10.1385/0-89603-335-X:111
Publisher Name: Humana Press
Print ISBN: 978-0-89603-335-1
Online ISBN: 978-1-59259-586-0
eBook Packages: Springer Protocols