Abstract Background Glioblastoma is among the most common primary malignancy with a poor medium survival post-diagnosis. The incredible heterogeneity of glioblastoma highlights its incurable nature. Methods to overcome difficulties leaded by glioblastoma heterogeneity remain to be explored. Here we present our Human2Human personalized autografted BRAF(V600E) mutant glioblastoma model, focusing on the idea of precision. This model, with an inoculation of self-derived glioblastoma cells, can imitate the tumor growth, invasion, metabolism and microenvironmental crosstalk within its “native” microenvironment and allows us to investigate the influence of different chemotherapies on the immunosuppressive tumor microenvironment from each specific individual glioblastoma patient. Material and Methods Non-neoplastic cortical tissue was obtained from a BRAF(V600E) mutant glioblastoma patient during surgical operation. Tissue was sectioned and inoculated with autografted glioblastoma cells in order to establish the ex-vivo Human2Human personalized brain slice model. Slice viability and tumor growth were monitored throughout the culture period, with and without day-wise refreshed treatments of clinically proved BRAF/MEK inhibitors. Sections were fixed and stained post cultivation. Pathway proteins p-ERK, p-Akt based on BRAF signaling along with markers (TMEM119, Iba-1, CD3, CD68, GFAP and NeuN) for major cell types in the environment were stained. Single Nuclei RNA Sequencing and Spatially Resolved Transcriptomics were applied. Results Tumor growth quantification over the culture period revealed different tumor reaction and tolerance towards various chemotherapies. The combination of Vemurafenib + Trametinib exhibited more efficient therapy response in comparison with either Dabrafenib + Trametinib or Encorafenib + Trametinib. Immunofluorescence and immunohistochemistry based quantification referring to neurons (NeuN), astrocytes (GFAP) and microglia/macrophage cells (Iba-1) suggested no toxic effects of the drug combinations on the tumor microenvironment. BRAF pathway proteins and immune cells showed various activation patterns upon different treatment combinations on an immunofluorescence base. Single Nuclei RNA Sequencing revealed the mesenchymal differentiation of BRAF mutant glioblastoma cells. Spatially Resolved Transcriptomics characterized tumor recurrence and suggested the therapy response accurately and visually. Conclusion The combination of Vemurafenib + Trametinib shows strategy to this specific BRAF mutant glioblastoma patient. And therefore, this Human2Human personalized model has a potential to provide in-depth information of the spatio-temporal tumor differentiation ex-vivo, correct inter-patient bias, and model therapy response in a very short time frame to provide drug testing results for clinical decision making.