Journal: International Journal of Molecular Sciences

Article Title: Transplantation of Embryonic Spinal Cord Derived Cells Helps to Prevent Muscle Atrophy after Peripheral Nerve Injury

doi: 10.3390/ijms18030511

Figure Lengend Snippet: In vitro characterization of cells isolated from E14.5 rat embryo spinal cords. ( A ) P0 cells represent the fetal cells that were directly isolated from embryos and used for transplantation on the same day. Most P0 cells were already of neuronal lineage as shown by their positive III-β-tubulin staining (red). In addition, some Nestin-positive (green) neural progenitor cells were observed. P2 cells represent the cells that were cultured two passages to obtain pure neural progenitor cells (NPCs). Almost all cells at that time were positive for Nestin (green) showing their progenitor cell identity. However, some already differentiated neurons were also seen (red III-β-tubulin staining). DAPI (4′,6-diamidino-2-phenylindole) staining is shown in blue; ( B ) On the 12th day of culturing, some P2 cells were also seeded onto coverslips with “Differentiating medium” for in vitro differentiation experiment. On the 6th day of differentiation, most of the cells had differentiated into GFAP (Glial fibrillary acidic protein)-positive astrocytes (red), with Rip-positive oligodendrocytes (green) and III-β-tubulin-positive neurons (red) also observed. The presence of several Nestin-positive cells (green) indicates undifferentiated immature cells. DAPI staining is shown in blue.

Article Snippet: The following primary antibodies were used: mouse anti-Nestin antibody (1:1000, BD Pharmingen) for undifferentiated NPCs; rabbit anti-glial fibrillary acidic protein (GFAP) (1:1000, Sigma-Aldrich) for astrocytes; mouse anti-Rip for oligodendrocytes (1:200, Merck-Millipore) and rabbit anti-β-III-tubulin (1:1000, Covance, Princeton, NJ, USA) for neurons.

Techniques: In Vitro, Isolation, Transplantation Assay, Staining, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Transplantation of Embryonic Spinal Cord Derived Cells Helps to Prevent Muscle Atrophy after Peripheral Nerve Injury

doi: 10.3390/ijms18030511

Figure Lengend Snippet: Surviving GFP (Green fluorescent protein)-expressing cells (green) can be seen on longitudinal sections of the musculocutaneous nerve taken from the 6-week time point. The left panels are representative for the P0 cell graft while the right panels for P2 cell graft. Grafted P0 fetal cells seem to have retained their neuronal identity, expressing NF200 (red) and NeuN (red), with some ChAT positive cells (red) detected as well. In contrast, grafted P2 NPCs have mostly differentiated into GFAP (red) expressing astrocytes. However, a few NeuN-positive cells (red) and NF200-positive axons (red) can also be seen even though their GFP expression is really low.

Article Snippet: The following primary antibodies were used: mouse anti-Nestin antibody (1:1000, BD Pharmingen) for undifferentiated NPCs; rabbit anti-glial fibrillary acidic protein (GFAP) (1:1000, Sigma-Aldrich) for astrocytes; mouse anti-Rip for oligodendrocytes (1:200, Merck-Millipore) and rabbit anti-β-III-tubulin (1:1000, Covance, Princeton, NJ, USA) for neurons.

Techniques: Expressing