Epithelial Cells Using Lingual Organoid Culture Benjamin Tiano - - PowerPoint PPT Presentation

Defining the Potency of Sox2-Positive Taste Epithelial Cells Using Lingual Organoid Culture

Benjamin Tiano

Barlow Lab-UC Denver ITTSD Conference 2018

What we know already

• Single LGR5+ cells generate

renewing lingual organoids

(Ren et al., 2014)

• Basal proliferative Sox2+

cells give rise to all types of taste bud cells (Ohmoto et al.,

2017)

• Sox2 is expressed in LGR5+

cells (Ohmoto et al., 2017)

• Sox2 deletion in lingual

progenitors disrupts taste bud renewal (Castillo-Azofeifa

et al., 2018)

Sox2 Suzuki, 2008 Ohmoto et al., 2017 Sox2 LGR5-EGFP/ KCNQ1 Sox2/LGR5-EGFP/ KCNQ1

* *

What we know already

• Single LGR5+ cells generate

renewing lingual organoids

(Ren et al., 2014)

• Basal proliferative Sox2+

cells give rise to all types of taste bud cells (Ohmoto et al.,

2017)

• Sox2 is expressed in LGR5+

cells (Ohmoto et al., 2017)

• Sox2 deletion in lingual

progenitors disrupts taste bud renewal (Castillo-Azofeifa

et al., 2018)

Dissociate into single cell suspension FACS sort based on GFP expression level Sox2-GFP Circumvallate Papilla Collect tongues and epithelia Low-GFP MedLo- GFP HiMed-GFP High-GFP

Do they grow?

Do different Sox2+ populations exhibit different growth potentials?

1000 cells/well 500 cells/well

LGR5 is differentially expressed by organoids from different Sox2 expression bins

Acknowledgements

Barlow Lab

Linda Barlow Dany Gaillard Erin Golden Jenn Scott Lauren Shechtman Kelly Zaccone