Figure 1.

Induction of spheroids from subcutaneous fat tissue of eGFP transgenic rat and their SCs differentiation. (a) Efficient induction of spheroids from adipose cells of eGFP transgenic rats. Upon exposure to the induction medium containing bFGF and EGF, adherent adipose tissue derived cells were merged to the spheroid and free floating ball-like structures were formed approximately at 6 d. (b) Characterization of the spheroid by immunofluorescence staining with anti-nestin antibody and BrdU labeling. Nestin-immunoreactive cells were negative at the periphery of the spheroid but confined to the inner part of the spheroid at 8 d induction. Most cells in the spheroid showed cell-proliferating capacity. (c) Immunofluorescence staining with anti-O4, anti-A2B4, and anti-S100β antibodies to confirm glial differentiation of the spheroid-forming cells in comparison with nSCs isolated from normal sciatic nerve. (d) Quantification of A2B5, O4, and S100β positive cells among total eGFP-expressing cells after immunofluorescence staining. (e) RT-PCR analysis for SCs specific genes expression. Both eGFP-SC^I and eGFP-SC^N exhibited similar gene expression profiles. eGFP-SCs^I: induced Schwann cells of eGFP transgenic rat; eGFP-SCs^N: naïve Schwann cells of eGFP transgenic rat.