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Isolation and Enrichment of Human Adipose-derived Stromal Cells for Enhanced Osteogenesis
Published on: January 12, 2015
[Experiment on inducing human periodontal ligament stem cells into adipose cells]
Hui-xia He1, Hong-chen Liu, Dong-sheng Wang
1Institute of Dental Research, General Hospital of Chinese PLA, Beijing 100853, China.
Objective:
To explore the capability of human periodontal ligament stem cells (PDLSCs) differentiating into adipose cells in vitro and to determine their changes in cell morphology, structure and function during differentiation.
Methods:
PDLSCs isolated by magnetic-activated cell selection were treated continuously with adipogenic medium for 21 d. Then the cell morphology, ultrastructure, adipose specific markers of low density lipoprotein (LPL) and peroxisome proliferator activated receptor-gamma (PPAR-gamma) were analyzed by inverted contrast microscope, trans mission electron microscope (TEM), flow cytometry, immunofluorescence, RT-PCR and Western blot, respectively. These adipose-like cells were also identified by oil red O staining to determine the formation of lipid droplet, and the non-induced cells were used as control.
Results:
After continuous induction, the treated cells differentiated into adipose-like cells with round shape, and large amount of lipid drop in cytoplasm. 96.54% of the PDLSCs were found to differentiate into adipose cells as showed by flow cytometry, the specific markers of LPL mRNA and PPAR-gamma mRNA, and oil red O staining, respectively. Further, PPAR-gamma protein was detected in the induced cells in a time-dependent manner.
Conclusion:
Human PDLSCs have the potential of differentiating into adipose cells under appropriate condition, and the differentiated cells exhibited characteristics of adipose cells both from cell morphology and from their functions.
