Abstract
The neural stem-cell marker nestin is expressed in hair follicle stem cells located in the bulge area which are termed hair-follicle-associated pluripotent (HAP) stem cells. HAP stem cells can differentiate into neurons, glia, keratinocytes, smooth muscle cells, and melanocytes in vitro. Subsequently, we demonstrated that HAP stem cells could affect nerve and spinal cord regeneration in mouse models. We subsequently demonstrated that HAP stem cells differentiated into beating cardiac muscle cells. The differentiation potential to cardiac muscle is greatest in the upper part of the mouse whisker follicle. The beat rate of the cardiac muscle cells differentiated from HAP stem cells was stimulated by isoproterenol and inhibited by propanolol. The addition of activin A, bone morphogenetic protein 4, and basic fibroblast growth factor, along with isoproternal, induced the cardiac muscle cells to form tissue sheets of beating heart muscle cells. Under hypoxic conditions, HAP stem cells differentiated into troponin-positive cardiac-muscle cells at a higher rate that under normoxic conditions. Hypoxia did not influence the differentiation to other cell types. This method is appropriate for future use with human hair follicles to produce hHAP stem cells in sufficient quantities for future heart, nerve, and spinal cord regeneration in the clinic.
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References
Li L, Mignone J, Yang M, Matic M, Penman S, Enikolopov G, Hoffman RM (2003) Nestin expression in hair follicle sheath progenitor cells. Proc Natl Acad Sci U S A 100:9958–9961
Amoh Y, Li L, Katsuoka K, Penman S, Hoffman RM (2005) Multipotent nestin-positive, keratin-negative hair-follicle-bulge stem cells can form neurons. Proc Natl Acad Sci U S A 102:5530–5534
Amoh Y, Li L, Katsuoka K, Hoffman RM (2008) Multipotent hair follicle stem cells promote repair of spinal cord injury and recovery of walking function. Cell Cycle 7:1865–1869
Yashiro M, Mii S, Aki R, Hamada Y, Arakawa N, Kawahara K, Hoffman RM, Amoh Y (2015) From hair to heart: nestin-expressing hair-follicle-associated pluripotent (HAP) stem cells differentiate to beating cardiac muscle cells. Cell Cycle 14:2362–2366
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 48:301–305
Liu F, Uchugonova A, Kimura H, Zhang C, Zhao M, Zhang L, Koenig K, Duong J, Aki R, Saito N, Mii S, Amoh Y, Katsuoka K, Hoffman RM (2011) The bulge area is the major hair follicle source of nestin-expressing pluripotent stem cells which can repair the spinal cord compared to the dermal papilla. Cell Cycle 10:830–839
Amoh Y, Li L, Campillo R, Kawahara K, Katsuoka K, Penman S, Hoffman RM (2005) Implanted hair follicle stem cells form Schwann cells that support repair of severed peripheral nerves. Proc Natl Acad Sci U S A 102:17734–17738
Mii S, Duong J, Tome Y, Uchugonova A, Liu F, Amoh Y, Saito N, Katsuoka K, 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 114:1674–1684
Kajiura S, Mii S, Aki R, Hamada Y, Arakawa N, Kawahara K, Li L, Katsuoka K, Hoffman RM, Amoh Y (2015) Cryopreservation of the hair follicle maintains pluripotency of nestin-expressing hair follicle-associated pluripotent stem cells. Tissue Eng Part C Methods 21:825–831
Yu H, Fang D, Kumar SM, Li L, Nguyen TK, Acs G, Herlyn M, Xu X (2006) Isolation of a novel population of multipotent adult stem cells from human hair follicles. Am J Pathol 168:1879–1888
Yu H, Kumar SM, Kossenkov AV, Showe L, Xu X (2010) Stem cells with neural crest characteristics derived from the bulge region of cultured human hair follicles. J Invest Dermatol 130:1227–1236
Yamazaki A, Yashiro M, Mii S, Aki R, Hamada Y, Arakawa N, Kawahara K, Hoffman RM, Amoh Y (2016) Isoproterenol directs hair follicle-associated pluripotent (HAP) stem cells to differentiate in vitro to cardiac muscle cells which can be induced to form beating heart muscle tissue sheets. Cell Cycle 15:760–765
Shirai K, Hamada Y, Arakawa N, Yamazaki A, Tohgi N, Aki R, Mii S, Hoffman RM, Amoh Y (2017) Hypoxia enhances differentiation of hair follicle-associated-pluripotent (HAP) stem cells to cardiac muscle cells. J Cell Biochem 118:554–558
Tohgi N, Obara K, Yashiro M, Hamada Y, Arakawa N, Mii S, Aki R, Hoffman RM, Amoh Y (2017) Human hair-follicle associated pluripotent (hHAP) stem cells differentiate to cardiac-muscle cells. Cell Cycle 16:95–99
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Hoffman, R.M., Amoh, Y. (2018). Beating Heart Cells from Hair-Follicle-Associated Pluripotent (HAP) Stem Cells. In: Singh, S., Rameshwar, P. (eds) Somatic Stem Cells. Methods in Molecular Biology, vol 1842. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8697-2_18
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DOI: https://doi.org/10.1007/978-1-4939-8697-2_18
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