Imunophenotyping of EnSCs: To detect surface antigens, cells were characterized by flow cytometry after passage three.
Adipogenic differentiation was induced in the third passage cells by plating the EnSCs at 2 x [10.sup.4] cells per [cm.sup.2], allowing the cells to reach confluence and then incubating for a further 48 hr.
Reverse Transcription-Polymerase Chain Reaction (RT-PCR): RT-PCR analysis was done to monitor the expression of activation of peroxisome proliferator activated receptor- PPAR during the programming of EnSCs into adipocytes cell lineage.
Characterization of isolated human EnSCs: Human EnSCs could be isolated easily by their adherence to plastic flask.
Analysis of adipocytes differentiation: After only 12 days of adipogenic induction, small lipid droplets (arrows in Figure 3) were observed within EnSCs treated with differentiation-promoting medium (Figure 3B) whereas there was no lipid droplet in non-treating group at day 12 (Figure 3A).
The EnSCs are new source of mesenchymal stem cells (21-25).
EnSCs should also contribute to the development of novel regenerative therapies for reconstruction of soft tissue defects after tumor resections, extensive deep burns and lipodystrophy.
It may be concluded that the EnSCs in the plastic and reconstructive surgical procedures for repairing soft tissues defects are more convenient than other sources of stem cells due to the following properties.