As part of my third semester curriculum (M.Sc) undertook this internship to characterize and study the stemness and differentiation behaviour of Brain Tumour Stem Cells (BTSCs). This included the passaging and treating of cells with various reagents to investigate the behavioural changes due to prolonged in-vitro incubation by immunocytochemical staining and also to investigate the influence of retinoic acid (RA), cyclic-Adenosinemonophosphate (cAMP) on the expansion of key neural markers by Western Blot Assay. I also investigated the presence of antibodies CD133, CD44 and CD31 by in-vivo FACS analysis. Extracted RNA from cells to check the expression of MGMT by Quantitative Real Time PCR.As part of Thesis research, we tried to improvise the viability of Brain tumour tissue for further analysis as against the traditional method of SCID-Mice inoculation for analyses. We tried to optimize the tissue slice size and other parameters for the MTT Assay, BrdU treatment etc. We also worked on obtaining live tissue slices using the Vibratome for better in-vitro observation. DNA content and protein analysis are a part of the project ( different staining like Propidium Iodide, Pico-green). In-vitro culture with different growth media (serum-free/with & with-out growth factors etc.)