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Isolation and characterization of hair follicle stem cells from Arbas Cashmere goat

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Abstract

In this study, highly purified hair follicle stem cells from Arbas Cashmere goat (gHFSCs) were isolated using enzyme digestion and adhesion to type IV collagen. The biological characteristics of the gHFSCs were identified by morphological observation, growth curve, markers assay and differentiation in vitro. The gHFSCs were in small cell size with typical cobblestone morphology, good adhesion and high refractive index. Immunocytochemistry staining showed the cells were expressing Krt15, Krt19, CD34, Itgβ1 and Krt14. Cell growth curve indicated that cultured gHFSCs had strong proliferation ability. Krt14 and CD34 were high expressed at the mRNA level, respectively, 39.68 and 24.37 times of the Cashmere goat keratinocytes, and krt15 expression was 5.62 times and itgβ1 expression was 1.81 times higher (p < 0.01). Western blot detected the expression of all the above markers. After osteogenic induction, the cells were positive for Von Kossa staining and expressed Osteocalcin. Sulfated proteoglycans in cartilaginous matrices were positively stained by Alcian blue after chondrogenic induction and COL2A1 was expressed. In myogenic induction, Hoechst 33342 staining evidenced cytoplasm fusion and positive expression of MyoG was detected by immunocytochemistry.

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References

  • 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 USA 102:5530–5534

    Article  CAS  Google Scholar 

  • Bajpai VK, Mistriotis P, Andreadis ST (2012) Clonal multipotency and effect of long-term in vitro expansion on differentiation potential of human hair follicle derived mesenchymal stem cells. Stem Cell Res 8:74–84

    Article  CAS  Google Scholar 

  • Bickenbach JR, Chism E (1998) Selection and extended growth of murine epidermal stem cells in culture. Exp Cell Res 244:184–195

    Article  CAS  Google Scholar 

  • Bose A, Teh M-T, Mackenzie IC, Waseem A (2013) Keratin k15 as a biomarker of epidermal stem cells. Int J Mol Sci 14:19385–19398

    Article  CAS  Google Scholar 

  • Commo S, Gaillard O, Bernard BA (2000) The human hair follicle contains two distinct K19 positive compartments in the outer root sheath: a unifying hypothesis for stem cell reservoir? Differentiation 66:157–164

    Article  CAS  Google Scholar 

  • Cotsarelis G, Sun T-T, Lavker RM (1990) Label-retaining cells reside in the bulge area of pilosebaceous unit: implications for follicular stem cells, hair cycle, and skin carcinogenesis. Cell 61:1329–1337

    Article  CAS  Google Scholar 

  • Ernst N, Yay A, Bíró T, Tiede S, Humphries M, Paus R, Kloepper JE (2013) β1 Integrin signaling maintains human epithelial progenitor cell survival in situ and controls proliferation, apoptosis and migration of their progeny. PLoS One 8:e84356

    Article  Google Scholar 

  • Guo Z, Draheim K, Lyle S (2011) Isolation and culture of adult epithelial stem cells from human skin. JoVE 49. doi:10.3791/2561

  • Jaks V, Kasper M, Toftgård R (2010) The hair follicle—a stem cell zoo. Exp Cell Res 316:1422–1428

    Article  CAS  Google Scholar 

  • Jones PH, Watt FM (1993) Separation of human epidermal stem cells from transit amplifying cells on the basis of differences in integrin function and expression. Cell 73:713–724

    Article  CAS  Google Scholar 

  • Kloepper JE, Tiede S, Brinckmann J, Reinhardt DP, Meyer W, Faessler R, Paus R (2008) Immunophenotyping of the human bulge region: the quest to define useful in situ markers for human epithelial hair follicle stem cells and their niche. Exp Dermatol 17:592–609

    Article  Google Scholar 

  • Liu X, Zhang P, Liu S (2006) Breed conservation and utilization of Inner Mongolia Arabs Cashmere goat China. Anim Husb Vet Med 32:34–36

    CAS  Google Scholar 

  • Liu JY, Peng HF, Gopinath S, Tian J, Andreadis ST (2010) Derivation of functional smooth muscle cells from multipotent human hair follicle mesenchymal stem cells. Tissue Eng Part A 16:2553–2564

    Article  CAS  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCt method. Methods 25:402–408

    Article  CAS  Google Scholar 

  • Mistriotis P, Andreadis ST (2013) Hair follicle: a novel source of multipotent stem cells for tissue engineering and regenerative medicine. Tissue Eng Part B Rev 19:265–278

    Article  CAS  Google Scholar 

  • Nowak JA, Fuchs E (2009) Isolation and culture of epithelial stem cells. Methods Mol Biol 482:215–232. doi:10.1007/978-1-59745-060-7_14

    Article  CAS  Google Scholar 

  • Ohyama M, Terunuma A, Tock CL, Radonovich MF, Pise-Masison CA, Hopping SB, Brady JN, Udey MC, Vogel JC (2006) Characterization and isolation of stem cell-enriched human hair follicle bulge cells. J Clin Investig 116:249

    Article  CAS  Google Scholar 

  • Poblet E, Jimenez F, Godinez J, Pascual-Martín A, Izeta A (2006) The immunohistochemical expression of CD34 in human hair follicles: a comparative study with the bulge marker CK15. Clin Exp Dermatol 31:807–812

    Article  CAS  Google Scholar 

  • Purba TS, Haslam IS, Poblet E, Jiménez F, Gandarillas A, Izeta A, Paus R (2014) Human epithelial hair follicle stem cells and their progeny: current state of knowledge, the widening gap in translational research and future challenges. BioEssays 36:513–525

    Article  CAS  Google Scholar 

  • Quan R, Zheng X, Ni Y, Xie S, Li C (2014) Culture and characterization of rat hair follicle stem cells. Cytotechnology 1–8. doi:10.1007/s10616-014-9807-z

  • Schneider MR, Schmidt-Ullrich R, Paus R (2009) The hair follicle as a dynamic miniorgan. Curr Biol 19:R132–R142

    Article  CAS  Google Scholar 

  • Vidal VP, Chaboissier MC, Lützkendorf S, Cotsarelis G, Mill P, Hui CC, Ortonne N, Ortonne JP, Schedl A (2005) Sox9 is essential for outer root sheath differentiation and the formation of the hair stem cell compartment. Curr Biol 15:1340–1351

    Article  CAS  Google Scholar 

  • Zhu B, Xu T, Zhang Z, Ta N, Gao X, Hui L, Guo X, Liu D (2014) Transcriptome sequencing reveals differences between anagen and telogen secondary hair follicle-derived dermal papilla cells of the Cashmere goat (Capra hircus). Physiol Genomics 46:104–111

    Article  Google Scholar 

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Acknowledgments

We are grateful to the Inner Mongolia YIWEI white Cashmere goat farm that provided us the goats for this study. This project was supported by the Generation of Induced Pluripotent Stem Cells from Cashmere Goat [Grant No. 2011JQ03].

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Authors and Affiliations

  1. Key Laboratory of Mammalian Reproductive Biology and Biotechnology Ministry of Education, Inner Mongolia University, Hohhot, 010070, Inner Mongolia, China

    Nimantana He, Zhenguo Dong, Shorgan Bou & Dongjun Liu

  2. College of Life Science, Inner Mongolia University, Hohhot, 010021, Inner Mongolia, China

    Li Tao & Sirguleng Zhao

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  1. Nimantana He
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  2. Zhenguo Dong
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  3. Li Tao
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  4. Sirguleng Zhao
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  5. Shorgan Bou
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  6. Dongjun Liu
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Correspondence to Nimantana He or Dongjun Liu.

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He, N., Dong, Z., Tao, L. et al. Isolation and characterization of hair follicle stem cells from Arbas Cashmere goat. Cytotechnology 68, 2579–2588 (2016). https://doi.org/10.1007/s10616-016-9981-2

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