FOXN1GFP/w reporter hESCs enable identification of integrin-a4, HLA-DR, and EpCAM as markers of human PSC-derived FOXN1 + thymic epithelial progenitors

Chew-Li Soh; Antonietta Giudice; Robert Alexander Jenny; David A Elliott; Tanya Hatzistavrou; Suzanne J Micallef; Korosh Kianizad; Natalie L Seach; Juan Carlos Zuniga-Pflucker; Ann P Chidgey; Alan O Trounson; Susie [Susan] K Nilsson; David Norman Haylock; Richard L Boyd; Andrew G Elefanty; Edouard G Stanley

doi:10.1016/j.stemcr.2014.04.009

FOXN1GFP/w reporter hESCs enable identification of integrin-a4, HLA-DR, and EpCAM as markers of human PSC-derived FOXN1 + thymic epithelial progenitors

Chew-Li Soh, Antonietta Giudice, Robert Alexander Jenny, David A Elliott, Tanya Hatzistavrou, Suzanne J Micallef, Korosh Kianizad, Natalie L Seach, Juan Carlos Zuniga-Pflucker, Ann P Chidgey, Alan O Trounson, Susie [Susan] K Nilsson, David Norman Haylock, Richard L Boyd, Andrew G Elefanty, Edouard G Stanley

Research output: Contribution to journal › Article › Research › peer-review

37 Citations (Scopus)

Abstract

Thymic epithelial cells (TECs) play a critical role in T cell maturation and tolerance induction. The generation of TECs from in vitro differentiation of human pluripotent stem cells (PSCs) provides a platform on which to study the mechanisms of this interaction and has implications for immune reconstitution. To facilitate analysis of PSC-derived TECs, we generated hESC reporter lines in which sequences encoding GFP were targeted to FOXN1, a gene required for TEC development. Using this FOXN1 (GFP/w) line as a readout, we developed a reproducible protocol for generating FOXN1-GFP(+) thymic endoderm cells. Transcriptional profiling and flow cytometry identified integrin-beta4 (ITGB4, CD104) and HLA-DR as markers that could be used in combination with EpCAM to selectively purify FOXN1(+) TEC progenitors from differentiating cultures of unmanipulated PSCs. Human FOXN1(+) TEC progenitors generated from PSCs facilitate the study of thymus biology and are a valuable resource for future applications in regenerative medicine.

Original language	English
Pages (from-to)	925 - 937
Number of pages	13
Journal	Stem Cell Reports
Volume	2
Issue number	6
DOIs	https://doi.org/10.1016/j.stemcr.2014.04.009
Publication status	Published - 2014

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10.1016/j.stemcr.2014.04.009

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Soh, C.-L., Giudice, A., Jenny, R. A., Elliott, D. A., Hatzistavrou, T., Micallef, S. J., Kianizad, K., Seach, N. L., Zuniga-Pflucker, J. C., Chidgey, A. P., Trounson, A. O., Nilsson, S. S. K., Haylock, D. N., Boyd, R. L., Elefanty, A. G., & Stanley, E. G. (2014). FOXN1GFP/w reporter hESCs enable identification of integrin-a4, HLA-DR, and EpCAM as markers of human PSC-derived FOXN1 + thymic epithelial progenitors. Stem Cell Reports, 2(6), 925 - 937. https://doi.org/10.1016/j.stemcr.2014.04.009

@article{407053608e284427b1aaad45714f9212,

title = "FOXN1GFP/w reporter hESCs enable identification of integrin-a4, HLA-DR, and EpCAM as markers of human PSC-derived FOXN1 + thymic epithelial progenitors",

abstract = "Thymic epithelial cells (TECs) play a critical role in T cell maturation and tolerance induction. The generation of TECs from in vitro differentiation of human pluripotent stem cells (PSCs) provides a platform on which to study the mechanisms of this interaction and has implications for immune reconstitution. To facilitate analysis of PSC-derived TECs, we generated hESC reporter lines in which sequences encoding GFP were targeted to FOXN1, a gene required for TEC development. Using this FOXN1 (GFP/w) line as a readout, we developed a reproducible protocol for generating FOXN1-GFP(+) thymic endoderm cells. Transcriptional profiling and flow cytometry identified integrin-beta4 (ITGB4, CD104) and HLA-DR as markers that could be used in combination with EpCAM to selectively purify FOXN1(+) TEC progenitors from differentiating cultures of unmanipulated PSCs. Human FOXN1(+) TEC progenitors generated from PSCs facilitate the study of thymus biology and are a valuable resource for future applications in regenerative medicine.",

author = "Chew-Li Soh and Antonietta Giudice and Jenny, \{Robert Alexander\} and Elliott, \{David A\} and Tanya Hatzistavrou and Micallef, \{Suzanne J\} and Korosh Kianizad and Seach, \{Natalie L\} and Zuniga-Pflucker, \{Juan Carlos\} and Chidgey, \{Ann P\} and Trounson, \{Alan O\} and Nilsson, \{Susie [Susan] K\} and Haylock, \{David Norman\} and Boyd, \{Richard L\} and Elefanty, \{Andrew G\} and Stanley, \{Edouard G\}",

year = "2014",

doi = "10.1016/j.stemcr.2014.04.009",

language = "English",

volume = "2",

pages = "925 -- 937",

journal = "Stem Cell Reports",

issn = "2213-6711",

publisher = "Cell Press",

number = "6",

}

Soh, C-L, Giudice, A, Jenny, RA, Elliott, DA, Hatzistavrou, T, Micallef, SJ, Kianizad, K, Seach, NL, Zuniga-Pflucker, JC, Chidgey, AP , Trounson, AO , Nilsson, SSK, Haylock, DN, Boyd, RL, Elefanty, AG & Stanley, EG 2014, 'FOXN1GFP/w reporter hESCs enable identification of integrin-a4, HLA-DR, and EpCAM as markers of human PSC-derived FOXN1 + thymic epithelial progenitors', Stem Cell Reports, vol. 2, no. 6, pp. 925 - 937. https://doi.org/10.1016/j.stemcr.2014.04.009

TY - JOUR

T1 - FOXN1GFP/w reporter hESCs enable identification of integrin-a4, HLA-DR, and EpCAM as markers of human PSC-derived FOXN1 + thymic epithelial progenitors

AU - Soh, Chew-Li

AU - Giudice, Antonietta

AU - Jenny, Robert Alexander

AU - Elliott, David A

AU - Hatzistavrou, Tanya

AU - Micallef, Suzanne J

AU - Kianizad, Korosh

AU - Seach, Natalie L

AU - Zuniga-Pflucker, Juan Carlos

AU - Chidgey, Ann P

AU - Trounson, Alan O

AU - Nilsson, Susie [Susan] K

AU - Haylock, David Norman

AU - Boyd, Richard L

AU - Elefanty, Andrew G

AU - Stanley, Edouard G

PY - 2014

Y1 - 2014

N2 - Thymic epithelial cells (TECs) play a critical role in T cell maturation and tolerance induction. The generation of TECs from in vitro differentiation of human pluripotent stem cells (PSCs) provides a platform on which to study the mechanisms of this interaction and has implications for immune reconstitution. To facilitate analysis of PSC-derived TECs, we generated hESC reporter lines in which sequences encoding GFP were targeted to FOXN1, a gene required for TEC development. Using this FOXN1 (GFP/w) line as a readout, we developed a reproducible protocol for generating FOXN1-GFP(+) thymic endoderm cells. Transcriptional profiling and flow cytometry identified integrin-beta4 (ITGB4, CD104) and HLA-DR as markers that could be used in combination with EpCAM to selectively purify FOXN1(+) TEC progenitors from differentiating cultures of unmanipulated PSCs. Human FOXN1(+) TEC progenitors generated from PSCs facilitate the study of thymus biology and are a valuable resource for future applications in regenerative medicine.

AB - Thymic epithelial cells (TECs) play a critical role in T cell maturation and tolerance induction. The generation of TECs from in vitro differentiation of human pluripotent stem cells (PSCs) provides a platform on which to study the mechanisms of this interaction and has implications for immune reconstitution. To facilitate analysis of PSC-derived TECs, we generated hESC reporter lines in which sequences encoding GFP were targeted to FOXN1, a gene required for TEC development. Using this FOXN1 (GFP/w) line as a readout, we developed a reproducible protocol for generating FOXN1-GFP(+) thymic endoderm cells. Transcriptional profiling and flow cytometry identified integrin-beta4 (ITGB4, CD104) and HLA-DR as markers that could be used in combination with EpCAM to selectively purify FOXN1(+) TEC progenitors from differentiating cultures of unmanipulated PSCs. Human FOXN1(+) TEC progenitors generated from PSCs facilitate the study of thymus biology and are a valuable resource for future applications in regenerative medicine.

UR - http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4050347/pdf/main.pdf

UR - https://www.scopus.com/pages/publications/84902281058

U2 - 10.1016/j.stemcr.2014.04.009

DO - 10.1016/j.stemcr.2014.04.009

M3 - Article

SN - 2213-6711

VL - 2

SP - 925

EP - 937

JO - Stem Cell Reports

JF - Stem Cell Reports

IS - 6

ER -

FOXN1GFP/w reporter hESCs enable identification of integrin-a4, HLA-DR, and EpCAM as markers of human PSC-derived FOXN1 + thymic epithelial progenitors

Abstract

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NHMRC Research Fellowship

Innovative stem cell-based strategies to establish immune tolerance and tissue repair.

Cite this

FOXN1GFP/w reporter hESCs enable identification of integrin-a4, HLA-DR, and EpCAM as markers of human PSC-derived FOXN1 + thymic epithelial progenitors

Abstract

UN SDGs

Access to Document

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Projects

NHMRC Research Fellowship

Innovative stem cell-based strategies to establish immune tolerance and tissue repair.

Cite this