Abstract
Access to complex in vitro models that recapitulate the unique markers and cell-cell interactions of the hair follicle is rather limited. Creation of scalable, affordable, and relevant in vitro systems which can provide predictive screens of cosmetic ingredients and therapeutic actives for hair health would be highly valued. In this study, we explore the features of the microfollicle, a human hair follicle organoid model based on the spatio-temporally defined co-culture of primary cells. The microfollicle provides a 3D differentiation platform for outer root sheath keratinocytes, dermal papilla fibroblasts, and melanocytes, via epidermal-mesenchymal-neuroectodermal cross-talk. For assay applications, microfollicle cultures were adapted to 96-well plates suitable for medium-throughput testing up to 21 days, and characterized for their spatial and lineage markers. The microfollicles showed hair-specific keratin expression in both early and late stages of cultivation. The gene expression profile of microfollicles was also compared with human clinical biopsy samples in response to the benchmark hair-growth compound, minoxidil. The gene expression changes in microfollicles showed up to 75% overlap with the corresponding gene expression signature observed in the clinical study. Based on our results, the cultivation of the microfollicle appears to be a practical tool for generating testable insights for hair follicle development and offers a complex model for pre-clinical substance testing.
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Acknowledgments
We would like to thank Mike Davis, Xingtao Wei, Rachel Adams, and Jay Tiesman for their valuable help with the microarray analysis. We would also like to thank Chris Kelling, Rudi Lurz, Reyk Horland, and Sarah Tremmel for their valuable contributions in this work.
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The technology for generating MFs is the subject of a patent from the Technische Universität Berlin that includes G.L. and R.L. as inventors. U.M., G.L., and R.L. are shareholders of TissUse GmbH, and U.M. is CSO of TissUse GmbH, Germany, which is commercializing the MFs. All other authors declare no competing interests. T.D., S.Y., and P.H. are employees of The Procter & Gamble Company.
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ESM 1
Viability of Microfollicles after Minoxidil (Mx) treatment. Microfollicle viability was measured via fluoresence using PrestoBlue assay at day 4 (D4) before Mx application and every 48 hours following day 8 (D8) during Mx treatment. Data are shown as mean ±SD. The mean comparison was performed using unpaired t-test with Welch’s correction. Statistical significance is shown for corresponding VC versus 1% Mx application with (**) when p<0.01 and (***) when p<0.0001. UTC-Untreated Control, VC-Vehicle Control, 1% Mx-1% minoxidil (n=5-6 MF per timepoint). MF metabolism during minoxidil treatment was examined in 2 donors with similar results. Donor used for microarray experiment is shown. (PNG 36 kb)
ESM 2
Primary antibodies (in 1:100 dilution) and secondary antibodies (in 1:200 dilution) used in immunofluorescence stainings are listed. (PNG 116 kb)
ESM 3
P values < 0.05 are highlighted in red, positive values are highlighted in yellow and negative values are highlighted in green. (XLSX 83 kb)
ESM 4
(MP4 4481 kb)
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Ataç, B., Kiss, F.M., Lam, T. et al. The microfollicle: a model of the human hair follicle for in vitro studies. In Vitro Cell.Dev.Biol.-Animal 56, 847–858 (2020). https://doi.org/10.1007/s11626-020-00513-x
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DOI: https://doi.org/10.1007/s11626-020-00513-x