Isolation of Highly Viable Thymic Epithelial Cells for Use in In Vitro and In Vivo Experiments

  • Mi-Jeong Kim
  • Thomas SerwoldEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1899)


Thymic epithelial cells (TECs) play multiple essential roles in T-cell development and the establishment of immune tolerance, but their isolation can be challenging, and their low viability upon isolation complicates downstream experiments. A method that allows TECs to be isolated easily and to survive afterward will be useful for elucidating key questions in TEC biology. Here, we demonstrate a simple method to isolate highly viable TECs. Primary TECs isolated using papain together with collagenase IV and DNase I survive and proliferate in vitro. Moreover, these primary TECs functionally engraft after intrathymic transplantation into recipient mice. Thus, the methods described herein will be useful for elucidating the roles of TECs and TEC subsets in T-cell development and immune tolerance.

Key words

Thymic epithelial cells Intrathymic transplantation Papain Thymus 



MJK was funded by a Mary K. Iacocca Foundation Research Fellowship. TS received funding from a seed grant from the Harvard Stem Cell Institute. The authors gratefully acknowledge the Joslin Diabetes Center Flow Cytometry Core supported by an NIH Diabetes Research Center grant (NIH award P30DK036836) and the Harvard Stem Cell Institute. The authors acknowledge the excellent support of the Joslin Animal Facility.


  1. 1.
    Anderson G, Jenkinson EJ (2001) Lymphostromal interactions in thymic development and function. Nat Rev Immunol 1:31–40CrossRefGoogle Scholar
  2. 2.
    Palmer E (2003) Negative selection–clearing out the bad apples from the T-cell repertoire. Nat Rev Immunol 5:383–391CrossRefGoogle Scholar
  3. 3.
    Steinmann GG, Klaus B, Müller-Hermelink HK (1985) The involution of the ageing human thymic epithelium is independent of puberty. A morphometric study. Scand J Immunol 22:563–575CrossRefGoogle Scholar
  4. 4.
    Hale JS, Boursalian TE, Turk GL, Fink PJ (2006) Thymic output in aged mice. Proc Natl Acad Sci U S A 103:8447–8452CrossRefGoogle Scholar
  5. 5.
    Sempowski GD, Gooding ME, Liao HX, Le PT, Haynes BF (2002) T cell receptor excision circle assessment of thymopoiesis in aging mice. Mol Immunol 38:841–848CrossRefGoogle Scholar
  6. 6.
    Chinn IK, Blackburn CC, Manley NR, Sempowski GD (2012) Changes in primary lymphoid organs with againg. Semin Immunol 24:309–320CrossRefGoogle Scholar
  7. 7.
    van Ewijk W, Wang B, Holländer G, Kawamoto H, Spanopoulou E, Ito M, Amagai T, Jian YF, Germeraad WTV, Chen WF, Katsura Y (1999) Thymic microenvironments, 3-D versus 2-D? Semin Immunol 11:57–64CrossRefGoogle Scholar
  8. 8.
    Berrih S, Savino W, Cohen S (1985) Extracellular matrix of the human thymus: immunofluorescence studies on frozen sections and cultured epithelila cells. J Histochem Cytochem 33:655–664CrossRefGoogle Scholar
  9. 9.
    Lannes-Vieira J, Dardenne M, Savino W (1991) Extracellular matrix components of the mouse thymus microenvironment: ontogenetic studies and modulation by glucocorticoid hormones. J Histochem Cytochem 39:1539–1546CrossRefGoogle Scholar
  10. 10.
    Huettner JE, Baughman RW (1986) Primary culture of identified neurons from the visual cortex of postnatal rats. J Neurosci 6:3044–3060CrossRefGoogle Scholar
  11. 11.
    Sarthy PV, Balkema GW (1981) Retrograde labeling and dissociation of mouse retinal ganglion cells. Neurosci Lett 25:205–208CrossRefGoogle Scholar
  12. 12.
    Bolzon BJ, Cheung DW (1989) Isolation and characterization of single vascular smooth muscle cells from spontaneously hypertensive rats. Hypertension 14:137–144CrossRefGoogle Scholar
  13. 13.
    Weekes TE (1974) The preparation of a viable suspension of epithelial cells from the rumen mucosa of cattle. Comp Biochem Physiol 49:407–413CrossRefGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Joslin Diabetes CenterBostonUSA

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