Identification of Biological Characteristics with Human Keloid Fibroblast-like Mesenchymal Stem Cells
|School||Zunyi Medical College,|
|Keywords||keloid mesenchymal tem cell cell culture differentiation|
Objective:To investigate whether there is keloid-derived mesenchymal stem cells in human keloid. Isolation, cultivation and identification of biological characteristics with human keloid-derived mesenchymal stem cslls(hKMSCs).Methods:The hKMSCs were isolated from human keliod with two-step enzymatic dissociation, selection by subculture. The surface antigen phenotype (CD29, CD34, CD44, CD45, CD90) and the cell cycle analysis was analyzed by flow cytometry.The expression of CK19 and vimentin was identified by Immunocytochemisty.Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of Oct4. Induced the cells to differentiate into osteogenic, chondrogenic and adipogenic lineage.Results:Mesenchymal stem cell could be isolated from human keloid, the shape of cell was uniform after being passaged.Flow cytometry analysis showed that 65.18±3.96% of cells were in the phase of G0/G1、6.92±1.82% in the phase of S and 27.90±2.19% in the phase of G2/M. Flow cytometry showed that these cells expressed the typical MSC marker CD29, CD44, and CD90, but they failed to express hematopoictic cell surface markers, such as CD34 and CD45.The result of immunocytochemisty showed Vimentin and Oct4 was expressed in the hKMSCS derived from human keloid was positive while CK19 from epithelial cell markers was negative.RT-PCR analysis revealed the expression of Oct4 from pluripotent stem cell markers was positive.After being cultured in inducing medium, the cells differentiated into osteogenic, chondrogenic and adipogenic lineage.Conclusions:Mesenchymal-like stem cells exists in human keloid, such stem cells may play an important role in the pathogenesis of keloid.