| Objective: To investigate the antigens expression of adipose-derived stem cells (ADSCs), as well as the effects of cell differentiated regulation on the repair and regeneration of skin injuries.Methods: The antigens expression of adult adipose-derived stemcell(ADSCs) were detected by immunohistochemical method. ADSCs were isolated, cultured and expanded in vitro respectively. The antigens expression of ADSCs were detected by two-steps immunocytochemistry. ADSCs were induced toward adipocytic lineage and osteogenic lineage differentiation, and were labeled with BrdU or Hochest33258. The confluenceAdult Human epidermal keracytocyte(HEKa) (2-4×105) were heat-shocked at 43℃ for 40 min and cooled for 1-2 hours at 37℃, then 1-2×105 labeled human ADSCs were added later and co-cultured with HEKa. The phenotype of ADSCs was detected, the percentage of phenotype conversion was calculated, and the factors and signaling pathways involved in the course were examined 3d after co-culture. The labeled rat ADSCs were intravenously injected into allogeneic full-thickness skin wounds rats, then the labeled cells and the phenotype of these cells were detetected 3d and 7d after allogeneic transplantation. Next, autospecific adipocyte were overjeted on burn wound of swine, the speed of wound healing and quality of healing in wounds treated with fat autografting were enhanced, indicating that ADSCs had a potential to accelerate the speed of healing and improvethe quality of healing of open burn wounds.Results: (1)ADSCs expressed markers of CD49 CD44 CD71 CD105 no CD34, CD106, CD45, CK19 and CK10. Basal layer expressed CK19 and CK14. Immunocytochemistry showed ADSCs did not express the hematopoietic markers CD34 and CD45, as well as HEKa markers CK19 and CK14. ADSCs could beinduced toward adipocytic lineage and osteogenic lineage differentiation. Both BrdU and Hochest33258 stained nucleus of ADSCs, and the percentage of BrdU labeling was about 74% and Hochest33258 about 90% (2) the cellular phenotype conversion of adipose derived stem cells (ADSCs) to epidermal cells. The positive rate of CK19 was 10.44%, 8.22%, 4.49%, and of CK14 was 12.46% 8.19% 4.37% in condition medium group, the condition medium with EGF group and 10%FBS DMEM with EGF group at 3 d, respectively, and 11.85% 9.96% 3.14%, and14.25%, 10.13%, 4.78.% at 7 d, respectively, all of them were greater than CK19 was 1.38% 1.7% at 3d 7d, and CK14 was 1.7%, 2.11% in control group at 3d, 7d, (p<0.01). (3)The cell cycle was G1: 35.04%, G2: 0.00%, S: 64.96% at control group, and G1: 67.25%, G2: 2.46%, S: 30.29% at condition medium group, suggeseted that the superior of homogenating rat skin makes ADSCs proliferating. When adult rADSCs were cocultured with heat-shocked HEKa., a subset of adult rADSCs differentiated into HEKa., and the percentage of differentiation was enhanced by EGF. CK14 7,CK19 in direct contact cocluture both increased higher than in direct contact cocluture without heat-shocked group (3) ①rADSCs the improve the speed of cellular wound model of HEKain vivo by make cell moving fasster,②The wound of HEKaby 5μl/ml 0.03% H2O2 or scrab 50-60cell area.and the amount of cells to wound was 8.2±0.15 个/H and8.5±0.22 个/H AFTER12hours, but the amount of cells to wound was 0.(3)rADSCs tranlant in vivo indicated that yhe area of experi,ental group was 59.6%, Ad VEGF165 infected rADSCs secrete VEGF mor 35% than control group.Conclusions: ADSCs derived mesenchymal stem cells has the potential and effect for wound healing and injuries repairing of skin injuries.
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