Figure 3.
Human MSC-CM100kD contains HGF and HGF promotes functional and histological recovery in EAE. A. Western blot of 3 samples of active MSC-CM100kD show the presence of HGF. B. Treatment with HGF (1 injection every other day for a total of 3 over a 5 day period, n=10) results in functional improvement compared to controls. Both dosage and route of delivery influence the efficacy of HGF to modulate disease. Intravenous (IV) injections are more effective than intraperitoneal (IP) injections. Arrows indicate treatment initiation day. C. HGF improves tissue histology. Sections from animals following 100ng IV injections at 30 days after immunization with MOG35–55. The improvement in myelination is apparent by Luxol Fast Blue staining. The reduction in immune cell infiltrates is evident in H&E and anti-CD3 labeled sections and confirmed in Toluidine Blue stained 1μm sections. Lower panel are representative electron micrographs through spinal cord lesion areas in EAE control and HGF treated animals 17 days after initiation of treatment. D. Upper Analysis of myelin thickness vs. axon diameter in lesion areas of control and HGF treated animals demonstrates thicker myelin in HGF treated animals compared to controls. Lower: Comparison of the relative axons diameters in lesion areas of control EAE and HGF treated animals demonstrates a reduction in small diameter fibers and an increase in medium diameter fibers in HGF treated animals. Bars = 500μm in C LFB (top), 50μm in C LFB (2ndpanel), H&E, CD3, Tol Blue and 2μm in the electron micrographs.