PHENOTYPIC CHANGES OF EPIDERMAL STEM CELLS DIFFERENTIATING INTO SWEAT GLANDS CELLS IN VITRO AND ITS MECHANISM_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

CHENFuhuan , YUEXiaotong ,  SONGHuifeng , CHAIJiake , HEXiuye , WANGTongmin , LIULingying

Department of Burn and Plastic Surgery, the First Affiliated Hospital of PLA General Hospital, Beijing, 100048, P. R. China;

Corresponding?author：

SONGHuifeng, Email: song_huifeng@126.com

Keywords：

Skin; Epidermal stem cells; Sweat gland cells; Cell differentiation; Phenotype; Signal pathway

DOI：

10.7507/1002-1892.20160049

Video：

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Objective To explore the phenotypic changes of epidermal stem cells (ESCs) differentiating into sweat glands cells (SGCs) in vitro and its mechanisms. Methods ESCs and SGCs were isolated and cultured in vitro, which were identified using immunofluorescence staining. ESCs at passage 2 were divided into 4 groups: ESCs and SGCs co-cultured by Transwell plates in group A, ESCs cultured by simply adding sweat supernatant in group B, ESCs and SGCs co-cultured on Transwell plate adding epidermal growth factor (EGF) (60 ng/mL) in group C, and ESCs and SGCs co-cultured on transwell plate adding PD98059 (10 mmol/L) in group D. The inverted microscope was used for observing the morphology of ESCs, flow cytometry for detecting ESCs positive phenotype, and Western blot for exploring mitogen-activated protein kinase/extracellular signal regulated kinase (MAPK/ERK) pathway. Results The morphology observation and immunofluorescence staining suggested that cultured cells were ESCs and SGCs. The inverted phase contrast microscope observation showed that cells had similar morphological changes, with flat polygonal shape at 9 days in groups A, C, and D; cells had slow morphological change in group B, and had similar change to that of other groups at 12 days. Significant decreasing of β₁-integrin expression and increasing of carcino-embryonic antigen (CEA) expression of ESCs were observed in group A when compared with group B, which was inhibited by EGF (group C) and enhanced by PD98059 (group D), and there were significant differences among groups A, C, and D (P<0.05). High level of ERK expression was displayed in 4 groups, but it was significantly lower in group B than the other 3 groups (P<0.05). The expression of phosphorylation ERK was the highest in group A and was the lowest in group C, showing significant difference among 4 groups (P<0.05). Conclusion ESCs can be induced to differentiate into SGCs with the phenotypic changes under the condition of co-cultured by Transwell plates. The MAPK/ERK pathway plays a key role in the differentiation of ESCs into SGCs.

Citation： CHENFuhuan, YUEXiaotong, SONGHuifeng, CHAIJiake, HEXiuye, WANGTongmin, LIULingying. PHENOTYPIC CHANGES OF EPIDERMAL STEM CELLS DIFFERENTIATING INTO SWEAT GLANDS CELLS IN VITRO AND ITS MECHANISM. Chinese Journal of Reparative and Reconstructive Surgery, 2016, 30(2): 245-250. doi: 10.7507/1002-1892.20160049 Copy

1.	?陳甫寰, 宋慧鋒, 王統民, 等. 汗腺細胞在人體皮膚創傷及修復中的研究進展. 中國美容醫學, 2015, 24(19):73-77.
2.	Boehnke K, Falkowska-Hansen B, Stark HJ, et al. Stem Cells of the human epidermis and their niche; composition and function in epidermal regeneration and carcinogenesis. Carcinogenesis, 2012, 33(7):1247-1258.
3.	Tao R, Sun TJ, Han YQ, et al. Epimorphin-induced differentiation of human umbilical cord mesenchymal stem cells into sweat gland cells. Eur Rev Med Pharmacol Sci, 2014, 18(9):1404-1410.
4.	Schneider MR, Werner S, Paus R, et al. The epidermal growth factor receptor and its ligands in skin biology and pathology. Am J Pathol, 2008, 173(1):14-24.
5.	Li HH, Zhou G, Fu XB, et al. Antigen expression of human eccrine sweat glands. J Cutan Pathol, 2009, 36(3):318-324.
6.	Charruyer A, Ghadially R. What's new in dermatology:epidermal stem cells. G Ital Dermatol Venereol, 2011, 146(1):57-67.
7.	Chuong CM, Randall VA, Widelitz RB, et al. Physiological regeneration of skin appendages and implications for regenerative medicine. Physiology (Bethesda), 2012, 27(2):61-72.
8.	Voog J, Jones DL. Stem cells and the niche:a dynamic duo. Cell Stem Cell, 2010, 6(2):103-115.
9.	李海紅, 付小兵, 周崗, 等. 人骨髓間充質干細胞與汗腺細胞共培養誘導細胞表型轉化的初步研究. 中華醫學雜志, 2005, 85(27):1885-1889.
10.	陳甫寰, 宋慧鋒, 岳曉彤, 等. 成人表皮干細胞和汗腺細胞的體外分離和培養. 中國美容醫學, 2015, 24(14):36-42.
11.	Kim JH, Kwon J, Lee HW, et al. Protein tyrosine kinase 7 plays a tumor role by inhibiting ERK and AKT phosphorylation in lung cancer. Oncol Rep, 2014, 31(6):2708-2712.
12.	Goldsmith EJ, Min X, He H, et al. Structural studies of MAPKinase cascade components. Methods Mol Biol, 2010, 661:223-237.
13.	Aznavoorine S, Stracke ML, Parsons J, et al. Interin mediates chemotactic and haptotatic motility in human melanoma cells through different signaling pathways. J Biol Chem, 1996, 271(6):3247-3254.
14.	Brakebusch C, F?ssler R. beta 1 integrin function in vivo:adhesion, migration and more. Cancer Metastasis Rev, 2005, 24(3):403-411.
15.	Stollery N. Sebaceous and sweat gland disorders. Practitioner, 2013, 257(1757):32-33.
16.	Joannides A, Gaughwin P, Scotc M, et al. Posmatal astrocytes promote neural induction from adult human bone marrow-derived stem cell. Hematother Stem Cell Res, 2003, 12(6):681-688.
17.	Kicic A, Shen WY, Wilson AS, et al. Differention of marrow stromal cells into photoreceptors in the rat eye. J Neurosci, 2003, 23(21):7742-7749.
18.	Rescan C, Le Bras S, Lefebvre VH, et al. EGF-induced proliferation of adult human pancreastic duct cell is mediated by MEK/ERK cascade. Lab Invest, 2005, 85(1):65-74.
19.	Yang W, Chen Y, Zhang Y, et al. Extracellular signal-regulated kinase 1/2 mitogen-activated protein kinase pathway is involved in myostain-regulated differentiateon repression. Cancer Res, 2006, 66(3):1320-1326.
20.	Shikiji T, Minami M, Inoue T, et al. Keratinocytes can differentiate into eccrine sweat ducts in vitro:involvement of epidermal growth factor and fetal bovine serum. J Dermatol Sci, 2003, 33(3):141-150.
21.	Pispa J, Pummila M, Barker PA, et al. Edar and Troy signaling pathways act redundantly to regulate initiation of hair follicle development. Hum Mol Genet, 2008, 17(21):3380-3391.
22.	Schmid K, Bago-Horvath Z, Berger W, et al. Dual inhibition of EGFR and mTOR Pathways in small cell lung cancer. Br J Cancer, 2010, 103(5):622-628.

1. ?陳甫寰, 宋慧鋒, 王統民, 等. 汗腺細胞在人體皮膚創傷及修復中的研究進展. 中國美容醫學, 2015, 24(19):73-77.
2. Boehnke K, Falkowska-Hansen B, Stark HJ, et al. Stem Cells of the human epidermis and their niche; composition and function in epidermal regeneration and carcinogenesis. Carcinogenesis, 2012, 33(7):1247-1258.
3. Tao R, Sun TJ, Han YQ, et al. Epimorphin-induced differentiation of human umbilical cord mesenchymal stem cells into sweat gland cells. Eur Rev Med Pharmacol Sci, 2014, 18(9):1404-1410.
4. Schneider MR, Werner S, Paus R, et al. The epidermal growth factor receptor and its ligands in skin biology and pathology. Am J Pathol, 2008, 173(1):14-24.
5. Li HH, Zhou G, Fu XB, et al. Antigen expression of human eccrine sweat glands. J Cutan Pathol, 2009, 36(3):318-324.
6. Charruyer A, Ghadially R. What's new in dermatology:epidermal stem cells. G Ital Dermatol Venereol, 2011, 146(1):57-67.
7. Chuong CM, Randall VA, Widelitz RB, et al. Physiological regeneration of skin appendages and implications for regenerative medicine. Physiology (Bethesda), 2012, 27(2):61-72.
8. Voog J, Jones DL. Stem cells and the niche:a dynamic duo. Cell Stem Cell, 2010, 6(2):103-115.
9. 李海紅, 付小兵, 周崗, 等. 人骨髓間充質干細胞與汗腺細胞共培養誘導細胞表型轉化的初步研究. 中華醫學雜志, 2005, 85(27):1885-1889.
10. 陳甫寰, 宋慧鋒, 岳曉彤, 等. 成人表皮干細胞和汗腺細胞的體外分離和培養. 中國美容醫學, 2015, 24(14):36-42.
11. Kim JH, Kwon J, Lee HW, et al. Protein tyrosine kinase 7 plays a tumor role by inhibiting ERK and AKT phosphorylation in lung cancer. Oncol Rep, 2014, 31(6):2708-2712.
12. Goldsmith EJ, Min X, He H, et al. Structural studies of MAPKinase cascade components. Methods Mol Biol, 2010, 661:223-237.
13. Aznavoorine S, Stracke ML, Parsons J, et al. Interin mediates chemotactic and haptotatic motility in human melanoma cells through different signaling pathways. J Biol Chem, 1996, 271(6):3247-3254.
14. Brakebusch C, F?ssler R. beta 1 integrin function in vivo:adhesion, migration and more. Cancer Metastasis Rev, 2005, 24(3):403-411.
15. Stollery N. Sebaceous and sweat gland disorders. Practitioner, 2013, 257(1757):32-33.
16. Joannides A, Gaughwin P, Scotc M, et al. Posmatal astrocytes promote neural induction from adult human bone marrow-derived stem cell. Hematother Stem Cell Res, 2003, 12(6):681-688.
17. Kicic A, Shen WY, Wilson AS, et al. Differention of marrow stromal cells into photoreceptors in the rat eye. J Neurosci, 2003, 23(21):7742-7749.
18. Rescan C, Le Bras S, Lefebvre VH, et al. EGF-induced proliferation of adult human pancreastic duct cell is mediated by MEK/ERK cascade. Lab Invest, 2005, 85(1):65-74.
19. Yang W, Chen Y, Zhang Y, et al. Extracellular signal-regulated kinase 1/2 mitogen-activated protein kinase pathway is involved in myostain-regulated differentiateon repression. Cancer Res, 2006, 66(3):1320-1326.
20. Shikiji T, Minami M, Inoue T, et al. Keratinocytes can differentiate into eccrine sweat ducts in vitro:involvement of epidermal growth factor and fetal bovine serum. J Dermatol Sci, 2003, 33(3):141-150.
21. Pispa J, Pummila M, Barker PA, et al. Edar and Troy signaling pathways act redundantly to regulate initiation of hair follicle development. Hum Mol Genet, 2008, 17(21):3380-3391.
22. Schmid K, Bago-Horvath Z, Berger W, et al. Dual inhibition of EGFR and mTOR Pathways in small cell lung cancer. Br J Cancer, 2010, 103(5):622-628.

Chinese Journal of Reparative and Reconstructive Surgery

PHENOTYPIC CHANGES OF EPIDERMAL STEM CELLS DIFFERENTIATING INTO SWEAT GLANDS CELLS IN VITRO AND ITS MECHANISM

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