CO-TRANSPLANTATION OF MOUSE EPIDERMIS AND DERMIS CELLS IN INDUCING HAIR FOLLICLE REGENERATION_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

CHENLin ^1,2 , XIJiafei ^1,2 , LIUDaqing ^1,2 , ZHANGXiuyuan ^1,2 , LÜYang ^1,2 , LIJing ^1,2 , WANGJingxue ^1,2 , ZHOUJunnian ^1,2 , NANXue ^1,2 ,  YUEWen ^1,2 ,  PEIXuetao ^1,2

1. Laboratory of Stem Cell and Regenerative Medicine, Institute of Field Transfusion Medicine, Academy of Military Medical Sciences, Beijing, 100850, P. R. China;
2. South China Research Center for Stem Cell & Regenerative Medicine, Academy of Military Medical Sciences;

Corresponding author：

YUEWen, Email: wenyue226@126.com; PEIXuetao, Email: peixt@nic.bmi.ac.cn

Keywords：

Hair follicle stem cells; Cell transplantation; Hair follicle regeneration; Mouse; Nude mouse

DOI：

10.7507/1002-1892.20160097

Video：

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Objective To investigate the co-transplantation of C57-green fluorescent protein (GFP) mouse epidermis and dermis cells subcutaneously to induce the hair follicle regeneration. Method C57-GFP mouse epidermis and dermis were harvested for isolation the mouse epidermis and dermis cells. The morphology of epidermis and dermis mixed cells at ratio of 1:1 of adult mouse, dermis cells of adult mouse, cultured 3rd generation dermis cells were observed by fluorescence microscope. Immunocytochemistry staining was used to detect hair follicle stem cells markers in cultured 3rd generation dermis cells from new born C57-GFP mouse. And then the epidermis and dermis mixed cells of adult mouse (group A), dermis cells of adult mouse (group B), cultured 3rd generation dermis cells of new born mouse (group C), and saline (group D) were transplanted subcutaneously into Balb/c nude mice. The skin surface of nude mice were observed at 4, 5, 6 weeks of transplantation and hair follicle formation were detected at 6 weeks by immunohistochemistry staining. Results The isolated C57-GFP mouse epidermis and dermis cells strongly expressed the GFP under the fluorescence microscope. Immunocytochemistry staining for hair follicle stem cells markers in cultured 3rd generation dermis cells showed strong expression of Vimentin and α-smooth muscle actin, indicating that the cells were dermal sheath cells; some cells expressed CD133, Versican, and cytokeratin 15. After transplanted for 4-6 weeks, the skin became black at the injection site in group A, indicating new hair follicle formation. However, no color change was observed in groups B, C, and D. Immunohistochemical staining showed that new complete hair follicles structures formed in group A. GFP expression could be only observed in the hair follicle dermal sheath and outer root sheath in group B, and it could also be observed in the hair follicle dermal sheath, outer root sheath, dermal papilla cells, and sweat gland in group C. The expression of GFP was negative in group D. Conclusions Co-transplantation of mouse epidermis and dermis cells can induce the hair follicle regeneration by means of interaction of each other. And transplantation of isolated dermis cells or cultured dermis cells individually only partly involved in the hair follicles formation.

Citation： CHENLin, XIJiafei, LIUDaqing, ZHANGXiuyuan, LÜYang, LIJing, WANGJingxue, ZHOUJunnian, NANXue, YUEWen, PEIXuetao. CO-TRANSPLANTATION OF MOUSE EPIDERMIS AND DERMIS CELLS IN INDUCING HAIR FOLLICLE REGENERATION. Chinese Journal of Reparative and Reconstructive Surgery, 2016, 30(4): 485-490. doi: 10.7507/1002-1892.20160097 Copy

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2.	Hagner A, Biernaskie J. Isolation and differentiation of hair follicle-derived dermal precursors. Methods Mol Biol, 2013, 989:247-263.
3.	Aoi N, Inoue K, Kato H, et al. Clinically applicable transplantation procedure of dermal papilla cells for hair follicle regeneration. J Tissue Eng Regen Med, 2012, 6(2) :85-95.
4.	Horne KA, Jahoda CA, Oliver RF. Whisker growth induced by implantation of cultured vibrissa dermal papilla cells in the adult rat. J Embryol Exp Morphol, 1986, 97:111-124.
5.	Oshimori N, Fuchs E. Paracrine TGF-β signaling counterbalances BMP-mediated repression in hair follicle stem cell activation. Cell Stem Cell, 2012, 10(1) :63-75.
6.	Dong L, Hao H, Liu J, et al. Wnt1a maintains characteristics of dermal papilla cells that induce mouse hair regeneration in a 3D preculture system. J Tissue Eng Regen Med, 2015.[Epub ahead of print].
7.	Zhang P, Kling RE, Ravuri SK, et al. A review of adipocyte lineage cells and dermal papilla cells in hair follicle regeneration. J Tissue Eng, 2014, 5:2041731414556850.
8.	Lichti U, Anders J, Yuspa SH. Isolation and short-term culture of primary kerationcytes, hair follicle populations and dermal cells from newborn mice and kerationocytes form adult mice for in vitro analysis and for grafting to immunodeficient mice. Nat Protoc, 2008, 3(5) :799-810.
9.	Aoi N, Inoue K, Kato H, et al. Clinically applicable transplantation procedure of dermal papilla cells for hair follicle regeneration. J Tissue Eng Regen Med, 2012, 6(2) :85-95.
10.	Mokos ZB, Mosler EL. Advances in a rapidly emerging field of hair follicle stem cell research. Coll Antropol, 2014, 38(1) :373-378.
11.	Jensen KM, Driskell RR, Watt FM. Assaying proliferation and differentiation capacity of stem cells using disaggregated adult mouse epidermis. Nat Protoc, 2010, 5(5) :898-911.
12.	金海兰, 张海明, 曹谊林. 毛囊干细胞及其分离与识别方法的研究进展. 中国美容医学, 2008, 17(8) :1250-1253.
13.	Fernandes KJ, McKenzie IA, Mill P, et al. A dermal niche for mutipotent adult skin-derived precursor cells. Nat Cell Biol, 2004, 6(11) :1082-1093.
14.	Zheng Y, Du X, Wang W, et al. Organogenesis from dissociated cells:generation of mature cycling hair follicles from skin-derived cells. J Invest Dermatol, 2005, 124(5) :867-876.
15.	Shimomura Y, Christiano AM. Biology and genetics of hair. Annu Rev Genomics Hum Gene, 2010, 11:109-132.
16.	McElwee KJ, Kissling S, Wenzel E, et al. Cultured peribulbar dermal sheath cells can induce hair follicle development and contribute to the dermal sheath and dermal papilla. J Invest Dermatol, 2003, 121(6) :1267-1275.
17.	Lichti U, Weinberg WC, Goodman L, et al. In vivo regulation of murine hair growth:insights from grafting defined cell populations onto nude mice. J Invest Dermatol, 1993, 101(1 Suppl):124S-129S.
18.	Aoi N, Inoue K, Chikanishi T, et al. 1α, 25-dihydroxyvitamin D3 modulates the hair-inductive capacity of dermal papilla cells:therapeutic potential for hair regeneration. Stem Cells Transl Med, 2012, 1(18) :615-626.
19.	Driskell RR, Juneja VR, Connelly JT, et al. Clonal growth of dermal papilla cells in hydrogels reveals intrinsic differences between Sox2-positive and -negative cells in vitro and in vivo. J Invest Dermatol, 2012, 132(4) :1084-1093.
20.	张群, 杨光辉, 丛笑倩, 等. 差速贴壁法分选人毛囊干细胞的研究. 中华实验外科杂志, 2006, 23(6) :755-758.

1. Morgan BA. The dermal papilla:an instructive niche for epithelial stem and progenitor cells in development and regeneration of the hair follicle. Cold Spring Harb Perspect Med, 2014, 4(7) :a015180.
2. Hagner A, Biernaskie J. Isolation and differentiation of hair follicle-derived dermal precursors. Methods Mol Biol, 2013, 989:247-263.
3. Aoi N, Inoue K, Kato H, et al. Clinically applicable transplantation procedure of dermal papilla cells for hair follicle regeneration. J Tissue Eng Regen Med, 2012, 6(2) :85-95.
4. Horne KA, Jahoda CA, Oliver RF. Whisker growth induced by implantation of cultured vibrissa dermal papilla cells in the adult rat. J Embryol Exp Morphol, 1986, 97:111-124.
5. Oshimori N, Fuchs E. Paracrine TGF-β signaling counterbalances BMP-mediated repression in hair follicle stem cell activation. Cell Stem Cell, 2012, 10(1) :63-75.
6. Dong L, Hao H, Liu J, et al. Wnt1a maintains characteristics of dermal papilla cells that induce mouse hair regeneration in a 3D preculture system. J Tissue Eng Regen Med, 2015.[Epub ahead of print].
7. Zhang P, Kling RE, Ravuri SK, et al. A review of adipocyte lineage cells and dermal papilla cells in hair follicle regeneration. J Tissue Eng, 2014, 5:2041731414556850.
8. Lichti U, Anders J, Yuspa SH. Isolation and short-term culture of primary kerationcytes, hair follicle populations and dermal cells from newborn mice and kerationocytes form adult mice for in vitro analysis and for grafting to immunodeficient mice. Nat Protoc, 2008, 3(5) :799-810.
9. Aoi N, Inoue K, Kato H, et al. Clinically applicable transplantation procedure of dermal papilla cells for hair follicle regeneration. J Tissue Eng Regen Med, 2012, 6(2) :85-95.
10. Mokos ZB, Mosler EL. Advances in a rapidly emerging field of hair follicle stem cell research. Coll Antropol, 2014, 38(1) :373-378.
11. Jensen KM, Driskell RR, Watt FM. Assaying proliferation and differentiation capacity of stem cells using disaggregated adult mouse epidermis. Nat Protoc, 2010, 5(5) :898-911.
12. 金海兰, 张海明, 曹谊林. 毛囊干细胞及其分离与识别方法的研究进展. 中国美容医学, 2008, 17(8) :1250-1253.
13. Fernandes KJ, McKenzie IA, Mill P, et al. A dermal niche for mutipotent adult skin-derived precursor cells. Nat Cell Biol, 2004, 6(11) :1082-1093.
14. Zheng Y, Du X, Wang W, et al. Organogenesis from dissociated cells:generation of mature cycling hair follicles from skin-derived cells. J Invest Dermatol, 2005, 124(5) :867-876.
15. Shimomura Y, Christiano AM. Biology and genetics of hair. Annu Rev Genomics Hum Gene, 2010, 11:109-132.
16. McElwee KJ, Kissling S, Wenzel E, et al. Cultured peribulbar dermal sheath cells can induce hair follicle development and contribute to the dermal sheath and dermal papilla. J Invest Dermatol, 2003, 121(6) :1267-1275.
17. Lichti U, Weinberg WC, Goodman L, et al. In vivo regulation of murine hair growth:insights from grafting defined cell populations onto nude mice. J Invest Dermatol, 1993, 101(1 Suppl):124S-129S.
18. Aoi N, Inoue K, Chikanishi T, et al. 1α, 25-dihydroxyvitamin D3 modulates the hair-inductive capacity of dermal papilla cells:therapeutic potential for hair regeneration. Stem Cells Transl Med, 2012, 1(18) :615-626.
19. Driskell RR, Juneja VR, Connelly JT, et al. Clonal growth of dermal papilla cells in hydrogels reveals intrinsic differences between Sox2-positive and -negative cells in vitro and in vivo. J Invest Dermatol, 2012, 132(4) :1084-1093.
20. 张群, 杨光辉, 丛笑倩, 等. 差速贴壁法分选人毛囊干细胞的研究. 中华实验外科杂志, 2006, 23(6) :755-758.

Chinese Journal of Reparative and Reconstructive Surgery

CO-TRANSPLANTATION OF MOUSE EPIDERMIS AND DERMIS CELLS IN INDUCING HAIR FOLLICLE REGENERATION

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