Primary culture of hPDSCs with modified enzymatic digestion ⁃ explants method

Objective To explore methods using modified tissue enzymatic separations for culturing primary hDP⁃ SCs in vitro and further identify the cells produced. Methods Primary hDPSCs were cultured using the modified tis⁃ sue enzymatic separation method, and cells were identified by morphology, cell surface markers, and differentiation po⁃ tential and evaluated using flow cytometry and growth curves. Results The hDPSCs were successfully isolated using the modified tissue enzymatic separation method. The morphology of these cells was similar to that of fibroblasts and mesenchymal stem cells, and the growth curve was "S" ⁃type. The results of cell phenotype analysis indicated that the cells were positive for surface markers of mesenchymal stem cells, including CD29, CD44, and CD90, and negative for markers of hematopoietic stem cells, including CD34, CD45, and CD106. The cells were capable of differentiating into multiple cell types. Conclusion The modified tissue enzymatic separation method can successful be used to culture primary hDPSCs in vitro.