| Objective:To explore the CD phenotypic, protein expression and pluripotent differentiation of human hypertrophic scar fibroblasts cultured in vitro. For future research of the mechanisms of scarring providing preliminary study data.Methods:Fibroblasts were isolated and cultured by digestion method with enzyme from human hypertrophic scar of1year. The cells morphology was observed by Inverted Microscope, and the growing state of the third passage was harvested by the cell counting meter of Vi-CELL. The cell surface marker CD105, CD14, CD73, CD34, CD44, CD45and CD90were identified by Flow Cytometry. The expression of CK19, Oct-4and vimentin were detected by immunocytochemistry, and the expression of a-SMA were detected by immunofluorescence. The potential of fibroblasts of the third passage to differentiate into adipogenic, osteogenic and chondrogenic lineages was assayed.Results:The primary passage fibroblast showed the shape of small tadpole on second day, and became gradually the spindle shaped or irregular polygon with a growing disposition of radial arrangement. The growth curve showed the cells grew slowly on the first and second day cultured, cells grew quickly in3days and reached platform stage on the sixth or seventh day. The fibroblast highly expressed mesenchymal stem cell surface markers-CD73, CD105, CD44, CD90, but not expressed hematopoietic stem cell surface markers-CD14, CD34, CD45by Flow cytometry. And positive expressed vimentin, Oct-4and negative expressed CK19by Immunocytochemistry, and positive expressed a-SMA (a-Smoothmuscleactin) by immunofluorescence. Multidirectional differentiation induction indicated that the third passage cells could differentiate into adipogenic, osteogenic and chondrogenic lineages.Conclusion:Human hypertrophic scar-derived fibroblast show the biology characteristics of Mesenchymal Stem Cells, and these biology characteristics may play a central role in hypertrophic scarring and wound heal. |
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