Deciphering the mechanism that enables skincancer to metastasize to the brain and inhibiting its spread by 80% JCI_Insight
For drug efficacy in intervention studies, B16-F10 cells were intracranially inoculated stereotactically to the striatum of 8- to 10-week-old C57BL/6 male mice. Three days following cells’ inoculation, mice were treated i.v. with 100 mg/kg bindarit or PBS QOD. On days 10 and 13 after melanoma cells’ inoculation, mice were euthanized and immediately perfused with 4% paraformaldehyde in PBS. Brains were then harvested for further immunostaining analysis.
Healthy human brain samples were collected by Thomas Hyde at the Lieber Institute for Brain Development as described under an approved IRB protocol, 90-M-0142.Tumor-bearing mice were anesthetized using ketamine and xylazine and perfused with 4% PFA in PBS. Mouse brains were resected, then incubated with 4% PFA for 4 hours, followed by 0.5 M of-sucrose overnight. Tissues were then embedded in O.C.T. on dry ice and stored at –80°C.For immunostaining of brain tumors, frozen O.C.T.