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Aim:To explore the effect ofnitric oxide rN0、on the adhesion、proliferation and migration of epidermal stem cells 1n vitro and 1n vivo Method:Human epidermal stem cells were 1solated and cultured with the modified method of rapid adhesion to collagen IV The harvested cells were 1dentified by the expression of epidermal stem cell markers,integrin[31 and keratin 1 9,through immunofluorescence staining with the presence of Nitric Oxide rN0、donor S-nitroso-N-acetvlpenicillamine(SNAP)at different concentrations(0,1,10,100,500IJtmol/L),the effect ofN0 on the migration of cultured epidermal stem cells was detected by 1n vitro wound healing model and Transwell assay .,respectively After that the effect ofNO on the adhesion and proliferation of epidermal stem cells was determined by routine methods By BrdU retention experiment and superficial partial-thickness scald model.we detect the migration effect of exogenous N0 on the migration of epidermal stem cells in mouse Results:(1)Under an inverted phase contrast microscope, cultured 5-9d small clone formas.the cell bodies are close together assuming”Cobblestone-like”growth; develop for 10-14 days,cell proliferation significantly speeds up,small clones grow-larger,and form mutual connections Into round.oval or 1rregular shaped clones.or 70%to 80%cell fusion Immunofluorescence assay showed that the 1solated cells can express CK19 and Integrin B1 r21 The scratch test conducted after 1 2,24 1ndicated that 1~100 gmol/L of SNAP can promote ESC migration.ofwhich 100 gmol/L SNAP migration rate reached a peak compared with 0 IJtmol/L sNAP respectively(48 8土2 7)%,(82 1土15.8)%,t value, respectively,8 34,5 10,P values less than O 01]Transwell experiments showed that the number of cells transferred to a membrane 1n the treatment of 100amol/L of SNAP 1s significantly larger than 0 mnol/L SNAP rP=O 00)The adhesion assay showed that with 1 OO.500 mol/L sNAP acting on ESC.cell absorbance value was significantly higher than of 0 timol/L of SNAP(P Values are equal to0.00)Proliferation experiments showed that with 1 00,500 timol/L of SNAP acting on ESC 24,48 h,the proliferative effect is more significant (P<0.05)(3)Scald for 65.C and 3s was indentified as injury condition for superficial partial-thickness scald model according to histological observation by electrical scald instrument with constant temperature and pressure;The results of staining and BrdU retention experiment showed that after injection of BrdU for 1 day,all cell showed varying degrees of positive expression of BrdU,and the expression of BrdU in the nucleus of skin decreased gradually over time,after six weeks,positive cells mainly distributed in the eminence of hair follicles;It is found in the body of ESC migration experiments that in the BrdU possitive cells of regenerated epidermis,L-arginine group,L-—NMMA group and saline group were significantly different,L-arginine group compared with L-NMMA group was significantly increased Conclusion:The human epidermal stem cell could be isolated successfully with the method we used;Exogenous nitric oxide could promote the migration and proliferation ofhuman epidermal stem cells andinhibitits adhesionfunctioninvitro;NO enhance significantly the migration of ESC by the adhesion and proliferation effect,so that NO ultimately promote wound healing after bumin vivo.