Effects Of Electric Fields On Migration Of Heet Injuried Epidermal Stem Cells

Objective We wonder whether the thermal damage factors can affect the migration behaviors of epidermal stem cells(Ep SCs) and how an EF will affect on heet injured Ep SCs.Methods First, we isolated EpSCs from SD rats skin by using a two-step tripsinization and differential velocity adhesion method to isolate and purify cells and cultured them in K-sfm medium,then using immunofluorescence technique to identify EpSCs surface markers?1-integrin and K19. Second, we explored the characteristics of nomal EpSCs and selected electric intensity and charging time for further study.Third, we established thermal damage model using P2-P3 EpSCs and examined whether the thermal damage factors can affect the migration behavior of EpSCs and how EFs affect on heet injured EpSCs.Results Using collagen IV to isolate and purify cells can successfully acquire Ep SCs, which are small and usually irregularly polygonal in shape, which have high nuclear-cytoplasmic ratio, and the cell membranes have strong refraction and array into acobblestone-like structure.The cultured EpSCs express stem cell markers ?1-integrin and K19 simultaneously. We found that EFs can induce Ep SCs migrating to the cathode, the larger the EF strength, the higher the directedness values and the longer the more apparent the directional migration.And the threshold voltage for EpSCs galvanotaxis was 50 mV/mm. EpSCs of themal injuredby 53? 30 s showed the same characteristics as nomal EpSCs in EFs,and they migrated faster than nomal EpSCs, but there was no obvious influence on migratory direction. The threshold voltage for heet injured EpSCs was 25 mV/mm.Conclusion The method of using a two-step tripsinization and differential velocity adhesion method to isolate and purify cells, and then culture them in K-sfm medium can successfully acquire Ep SCs which we need. EFs can induce a slight themal injured Ep SCs to migrate to the cathode in a voltage- and time- dependent manner, and themal injured EpSCs migrate faster than nomal EpSCs. Heet injured Ep SCs increased sensitivity to EF stimulation and the threshold voltage decreased to 25 mV/mm. These results provide novel insights into our understanding on the mechanism of burn wound healing. Clinically,improving the galvanotactic response of cells might be an effective method to accelerate wound healing.