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. 2022 Aug;31(8):1276-1280.
doi: 10.1111/exd.14626. Epub 2022 Jun 23.

HaCaT cells as a model system to study primary cilia in keratinocytes

Gabriela Blanchard  1 , Christine Pich  1 , Daniel Hohl  1   2
Affiliations

HaCaT cells as a model system to study primary cilia in keratinocytes

Gabriela Blanchard et al. Exp Dermatol. 2022 Aug.

Abstract

Primary cilium (PC) is a microtubule-based organelle found on the apical surface of most mammalian cell types, playing a role in development and tissue homeostasis. Ciliopathies are a rapidly growing group of human diseases characterized by disordered cilium. PC plays an important role in pathogenesis of basal cell cancer, the most common human malignancy. A significant increase in ciliation has been observed in the epidermis of atopic dermatitis and psoriasis patients. Spontaneously immortalized human keratinocytes, HaCaT are a model to study the epidermal homeostasis and pathophysiology. In contrast to what has been previously described, here, we show that HaCaT can be efficiently ciliated. In HaCaT cells, differentiation significantly increased the number of ciliated cells and we were able to analyse in detail the ciliary length progression with duration of differentiation. As the number of recognized ciliopathies continues to increase, the importance of ciliary models also rises. Even though keratinocytes do not become as highly and rapidly ciliated as cell lines frequently used in ciliary studies, they are a better model for the study of skin ciliopathies. Detailed progression of ciliation in HaCaT could serve as the basis for ciliary studies in this cell line.

Keywords: cancer; differentiation; keratinocyte biology; keratinocytes; primary cilium.

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Conflict of interest statement

The authors declare no competing interests.

Figures

FIGURE 1
FIGURE 1
Differentiation of HaCaT keratinocytes increases their ciliation. (A) Western blot showing a representative image of K10 protein expression during differentiation of HaCaT. α‐tubulin used as a loading control. (B) Representative immunofluorescence images of acetylated tubulin (green) and rootletin (red) co‐staining in non‐differentiated and 7 days‐differentiated HaCaT. Scale bar, 10 μm. (C) Quantification of ciliated HaCaT upon non‐differentiated and 7 days differentiated conditions. Unpaired t test with Welch's correction (N = 10, 16). (D) Cilium length measured in HaCaT at D3, D5 and D7 of differentiation. Ordinary one‐way ANOVA test with Holm‐Šidák's correction (N = 64, 68, 59). (E) Representative images of SMO IN/OUT assay in 7 days‐differentiated HaCaT. Scale bar, 5 μm. Ac‐tubul, acectylated tubulin; IB, immunoblot; ND, non‐differentiated, D3, D5, D7: day 3, 5 and 7 of differentiation, respectively
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References

    1. Goetz SC, Anderson KV. The primary cilium: a signalling Centre during vertebrate development. Nat Rev Genet. 2010;11(5):331‐344. doi:10.1038/nrg2774 - DOI - PMC - PubMed
    1. Singla V, Reiter JF. The primary cilium as the cell's antenna: signaling at a sensory organelle. Science. 2006;313(5787):629‐633. doi:10.1126/science.1124534 - DOI - PubMed
    1. Verhey KJ, Gaertig J. The tubulin code. Cell Cycle. 2007;6(17):2152‐2160. doi:10.4161/cc.6.17.4633 - DOI - PubMed
    1. Yu I, Garnham CP, Roll‐Mecak A. Writing and reading the tubulin code. J Biol Chem. 2015;290(28):17163‐17172. doi:10.1074/jbc.R115.637447 - DOI - PMC - PubMed
    1. Malicki JJ, Johnson CA. The cilium: cellular antenna and central processing unit. Trends Cell Biol. 2017;27(2):126‐140. doi:10.1016/j.tcb.2016.08.002 - DOI - PMC - PubMed

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