Evaluation of TgH(CX3CR1-EGFP) mice implanted with mCherry-GL261 cells as an in vivo model for morphometrical analysis of glioma-microglia interaction

摘要: Glioblastoma multiforme is the most aggressive brain tumor. Microglia are prominent cells within glioma tissue and play important roles in tumor biology. This work presents an animal model designed for study of microglial cell morphology situ during gliomagenesis. It also allows a quantitative morphometrical analysis their activation by cells. The associates following types: 1- mCherry red fluorescent GL261 and; 2- EGFP microglia, present TgH(CX3CR1-EGFP) mouse line. First, mCherry-GL261 were implanted cortex mice. Epifluorescence − and confocal laser-scanning microscopy employed fixed sections, whereas two-photon (2P-LSM) was used to track microglia living animals. Implanted successfully developed tumors. They mimic behavior found human disease, with rapid increase size presence secondary tumors apart from injection site. As grows, progressively lost original shape, adopting heterogeneous diffuse at 14–18 d. Soma increased 10–52 μm. At this point, we focused on kinetics access tissues. 2P-LSM revealed intense microgliosis areas already shortly after implantation, i.e. 30 min. By microscopy, clusters around mass first 3 days. Then infiltrated area, where they remained all time points studied, 6–18 days. contact changes morphology, ramified amoeboid shape. Cell bodies enlarged 366 ± 0.0 μm2, quiescent 1310 ± 146.0 μm2, processes became shortened. GL261/CX3CR1 reported here valuable tool imaging growth, either sections or Remarkable advantages use immunocompetent animals simplified method without need immunohistochemical procedures.