Transient focal cerebral ischaemia was induced by middle cerebral artery occlusion (MCAo) for 30 mins, based on our previously published protocol using the intraluminal filament model.9 (link)
Briefly, a midline incision was made on the ventral surface of the neck and the left common carotid artery isolated and ligated. Topical anaesthetic (EMLA, 5% prilocaine and lidocaine, AstraZeneca, UK) was applied to skin incision sites prior to incision. The internal carotid artery and the pterygopalatine artery were temporarily ligated. A 6-0 monofilament (Doccol, Sharon, MA, USA) was introduced into the internal carotid artery via an incision in the common carotid artery. The filament was advanced approximately 10 mm into the common carotid with the filament making its way distal to the carotid bifurcation, beyond the origin of the middle cerebral artery. A 10 mm mark was made on the filament to visualise the required length to be inserted beyond the carotid bifurcation. At 3 h from start of occlusion, animals were treated under isoflurane anaesthesia with vehicle (serum-free Mesenpro media), 9.1 × 104 IL-1α conditioned MSCs or 9.1 × 104 non-conditioned MSCs by intra-arterial infusion with a syringe driver (20 µl cell suspension, 0.5 µl/sec), via the filament incision site. Animals were recovered and returned to normal housing prior to behavioural testing.
Briefly, a midline incision was made on the ventral surface of the neck and the left common carotid artery isolated and ligated. Topical anaesthetic (EMLA, 5% prilocaine and lidocaine, AstraZeneca, UK) was applied to skin incision sites prior to incision. The internal carotid artery and the pterygopalatine artery were temporarily ligated. A 6-0 monofilament (Doccol, Sharon, MA, USA) was introduced into the internal carotid artery via an incision in the common carotid artery. The filament was advanced approximately 10 mm into the common carotid with the filament making its way distal to the carotid bifurcation, beyond the origin of the middle cerebral artery. A 10 mm mark was made on the filament to visualise the required length to be inserted beyond the carotid bifurcation. At 3 h from start of occlusion, animals were treated under isoflurane anaesthesia with vehicle (serum-free Mesenpro media), 9.1 × 104 IL-1α conditioned MSCs or 9.1 × 104 non-conditioned MSCs by intra-arterial infusion with a syringe driver (20 µl cell suspension, 0.5 µl/sec), via the filament incision site. Animals were recovered and returned to normal housing prior to behavioural testing.
