Hair-Shaft Growth in Gelfoam® Histoculture of Skin and Isolated Hair Follicles

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1760)

Abstract

Human scalp skin with abundant hair follicles in various stages of the hair growth cycle was histocultured for up to 40 days on Gelfoam® at the air/liquid interface. The anagen hair follicles within the histoculture scalp skin produced growing hair shafts. Hair follicles could continue their cycle in histoculture; for example, apparent spontaneous catagen induction was observed both histologically and by the actual regression of the hair follicle. In addition, vellus follicles were shown to be viable at day 40 after initiation of culture. Follicle keratinocytes continued to incorporate [3H]thymidine for up to several weeks after shaft elongation had ceased. Intensive hair growth was observed in the pieces of shaved mouse skin histocultured on Gelfoam®. Isolated human and mouse hair follicles also produced growing hair shafts. By day 63 in histoculture of mouse hair follicles, the number of hair follicle-associated pluripotent (HAP) stem cells increased significantly and the follicles were intact. Gelfoam® histoculture of skin demonstrated that the hair follicle cells are the most sensitive to doxorubicin which prevented hair growth, thereby mimicking chemotherapy-induced alopecia in Gelfoam® histoculture.

Key words

Hair follicles, Chemotherapy Hair growth Human scalp Mouse Gelfoam® histoculture Hair follicle-associate-pluripotent (HAP) stem cells 

References

  1. 1.
    Strangeways DH (1931) The growth of hair in vitro. Arch Exp Zellforsch 11:344Google Scholar
  2. 2.
    Murray MR (1933) Development of the hair follicle and hair in vitro. Anat Rec 57(Supplement):74Google Scholar
  3. 3.
    Hardy MH (1951) The development of pelage hairs and vibrissae from skin in tissue culture. Ann N Y Acad Sci 53:546–561CrossRefPubMedGoogle Scholar
  4. 4.
    Frater R, Whitmore PG (1973) The in-vitro growth of post-embryonic hair. J Invest Dermatol 61:72–81CrossRefPubMedGoogle Scholar
  5. 5.
    Philpott M, Green M, Kealey T (1989) Studies on the biochemistry and morphology of freshly isolated and maintained rat hair follicles. J Cell Sci 93:409–418PubMedGoogle Scholar
  6. 6.
    Philpott M, Green M, Kealey T (1990) Human hair growth in vitro. J Cell Sci 97:463–471PubMedGoogle Scholar
  7. 7.
    Li L, Paus R, Margolis LB, Hoffman RM (1992) Hair shaft elongation, follicle growth, and spontaneous regression in long-term, gelatin sponge-supported histoculture of human scalp skin. Proc Natl Acad Sci U S A 89:8764–8768CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Kido T, Horigome T, Uda M, Adachi N, Hirai Y (2017) Generation of iPS-derived model cells for analyses of hair shaft differentiation. BioTechniques 63:131–134CrossRefPubMedGoogle Scholar
  9. 9.
    Ojeh N, Akgül B, Tomic-Canic M, Philpott M, Navsaria H (2017) In vitro skin models to study epithelial regeneration from the hair follicle. PLoS One 12:e0174389CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Su YS, Fan ZX, Xiao SE, Lin BJ, Miao Y, Hu ZQ, Liu H (2017) Icariin promotes mouse hair follicle growth by increasing insulin-like growth factor 1 expression in dermal papillary cells. Clin Exp Dermatol 42:287–294CrossRefPubMedGoogle Scholar
  11. 11.
    Chacón-Martínez CA, Klose M, Niemann C, Glauche I, Wickström SA (2017) Hair follicle stem cell cultures reveal self-organizing plasticity of stem cells and their progeny. EMBO J 36:151–164CrossRefPubMedGoogle Scholar
  12. 12.
    Zhou L, Yang K, Xu M, Andl T, Millar SE, Boyce S, Zhang Y (2016) Activating β-catenin signaling in CD133-positive dermal papilla cells increases hair inductivity. FEBS J 283:2823–2835CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Hwang I, Choi KA, Park HS, Jeong H, Kim JO, Seol KC, Kwon HJ, Park IH, Hong S (2016) Neural stem cells restore hair growth through activation of the hair follicle niche. Cell Transplant 25:1439–1451CrossRefPubMedGoogle Scholar
  14. 14.
    Paus R, Burgoa I, Platt CI, Griffiths T, Poblet E, Izeta A (2016) Biology of the eyelash hair follicle: an enigma in plain sight. Br J Dermatol 174:741–752CrossRefPubMedGoogle Scholar
  15. 15.
    Langan EA, Philpott MP, Kloepper JE, Paus R (2015) Human hair follicle organ culture: theory, application and perspectives. Exp Dermatol 24:903–911CrossRefPubMedGoogle Scholar
  16. 16.
    Sohn KM, Jeong KH, Kim JE, Park YM, Kang H (2015) Hair growth-promotion effects of different alternating current parameter settings are mediated by the activation of Wnt/β-catenin and MAPK pathway. Exp Dermatol 24:958–963CrossRefPubMedGoogle Scholar
  17. 17.
    Kiso M, Hamazaki TS, Itoh M, Kikuchi S, Nakagawa H, Okochi H (2015) Synergistic effect of PDGF and FGF2 for cell proliferation and hair inductive activity in murine vibrissal dermal papilla in vitro. J Dermatol Sci 79:110–118CrossRefPubMedGoogle Scholar
  18. 18.
    Yang XY, Jin K, Ma R, Yang JM, Luo WW, Han Z, Cong N, Ren DD, Chi FL (2015) Role of the planar cell polarity pathway in regulating ectopic hair cell-like cells induced by Math1 and testosterone treatment. Brain Res 1615:22–30CrossRefPubMedGoogle Scholar
  19. 19.
    Li L, Margolis LB, Hoffman RM (1991) Skin toxicity determined in vitro by three-dimensional, native-state histoculture. Proc Natl Acad Sci U S A 88:1908–1912CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Li L, Paus R, Margolis LB, Hoffman RM (1992) Hair growth in vitro from histocultured skin. In Vitro Cell Dev Biol 28A:479–481CrossRefPubMedGoogle Scholar
  21. 21.
    Li L, Paus R, Slominski A, Hoffman RM (1992) Skin histoculture assay for studying the hair cycle. In Vitro Cell Dev Biol 28A:695–698CrossRefPubMedGoogle Scholar
  22. 22.
    Mii S, Duong J, Tome Y, Uchugonova A, Liu F, Amoh Y, Hoffman RM (2013) The role of hair follicle nestin-expressing stem cells during whisker sensory-nerve growth in long-term 3D culture. J Cell Biochem 114:1674–1684CrossRefPubMedGoogle Scholar
  23. 23.
    Cao W, Li L, Mii S, Amoh Y, Liu F, Hoffman RM (2015) Extensive hair shaft elongation by isolated mouse whisker follicles in very long-term Gelfoam® histoculture. PLoS One 10:e0138005CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Uchugonova A, Zhao M, Weinigel M, Zhang Y, Bouvet M, Hoffman RM, Koenig K (2013) Multiphoton tomography visualizes collagen fibers in the tumor microenvironment that maintain cancer-cell anchorage and shape. J Cell Biochem 114:99–102CrossRefPubMedGoogle Scholar
  25. 25.
    Jahoda CAB, Home KA, Oliver RF (1984) Induction of hair growth by implantation of cultured dermal papilla cells. Nature 311:560–562CrossRefPubMedGoogle Scholar
  26. 26.
    Rogers G, Martinet N, Steinert P, Wynn P, Roop D, Kilkenny A, Morgan D, Yuspa SH (1987) Cultivation of murine hair follicles as organoids in a collagen matrix. J Invest Dermatol 89:369–379CrossRefPubMedGoogle Scholar
  27. 27.
    Paus R, Stenn KS, Link RE (1989) The induction of anagen hair growth in telogen mouse skin by cyclosporine A administration. Lab Investig 60:365–369PubMedGoogle Scholar
  28. 28.
    Paus R, Stenn KS, Link RE (1990) Telogen skin contains an inhibitor of hair growth. Br J Dermatol 122:777–784CrossRefPubMedGoogle Scholar
  29. 29.
    Paus R, Stenn KS, Elgjo K (1991) The epidermal pentapeptide pyroGlu-Glu-Asp-Ser-GlyOH inhibits murine hair growth in vivo and in vitro. Dermatologica 183:173–178CrossRefPubMedGoogle Scholar
  30. 30.
    Duong J, Mii S, Uchugonova A, Liu F, Moossa AR, Hoffman RM (2012) Real-time confocal imaging of trafficking of nestin-expressing multipotent stem cells in mouse whiskers in long-term 3-D histoculture. In Vitro Cell Dev Biol Anim 48:301–305CrossRefPubMedGoogle Scholar
  31. 31.
    Goldberg MT, Tackaberry LE, Hardy MH, Noseworthy JH (1990) Nuclear aberrations in hair follicle cells of patients receiving cyclophosphamide. A possible in vivo assay for human exposure to genotoxic agents. Arch Toxicol 64:116–121CrossRefPubMedGoogle Scholar
  32. 32.
    Jo SJ, Choi S-J, Yoon S-Y, Lee JY, Park W-S, Park P-J et al (2013) Valproic acid promotes human hair growth in in vitro culture model. J Dermatol Sci 72:16–24CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.AntiCancer Inc.San DiegoUSA
  2. 2.Department of SurgeryUniversity of CaliforniaSan DiegoUSA
  3. 3.Department of AnatomySecond Military Medical UniversityShanghaiChina

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