The development of xenograft glioblastoma implants in nude mice brain

Abstract

The inefficacity of the actual therapies for glioblastoma multiformis stimulates the researchers to search for new and innovative therapies. Therefore, the development of in vivo model for glioblastoma is an essential step during these researches, being a link between cells cultures studies and the first phases of clinical trials. In this paper, we present several procedures which have been performed for the first time in our country, such as: the cultivation and manipulation of U87MG line, the manipulation of athymic knock-out mice (NUDE Crl: CD-1 Foxn 1), the stereotactic inoculation of glioblastoma cells and finally the development of glioblastoma xenograft in the brain of inoculated nude mice. These results, which offer to the researchers from our country an in vivo model for glioblastoma, could be the start point for several projects oriented to the development of new therapies in glioblastoma, and could raise the performance of our scientific research to the European level.

Fig. 1

Microscopical aspects of Glioblastoma line U 87(Inverted microscope, X20)

Fig. 2

U 87 cells transfected with GFP–visualized at fluorescence microscope

Fig. 3

The phenotype of the nude mice used in our experiments

Fig. 4

Micro–instruments used for surgical experiments

Fig. 5

Micro–motor TC–Motor 3000

Fig. 6

Stereotaxic system: TAXIC–600 – WPI Stereotaxic Frame 18 deg. Ear Bars and UMP 3–1 injection system, (World Precision Instruments, Germany).

Fig. 7

The main anatomical landmarks

Fig. 8

Inoculation of trypane blue in order to demonstrate the site of inoculation–coronal section of the mouse brain

Fig. 9

Mouse post–inoculation–aspect of scalp incision

Fig. 10

Macroscopic (10A) and microscopic (10 B–inverted image) aspect of the glioblastoma xenograft at 7 days postinoculation (arrow). Colored with hematoxilin–eosin.

Fig. 11

The fluorescence microscopic aspect of the GFP transfected glioblastoma cells at 3 days post–inoculation. Computerized system acquisition has been used for this image

Fig. 12

Tumor characterized by hyper–cellurality. Image taken at 7 days post inoculation (Colored with DAPI).

Fig. 13

Multinucleate gigantic cells (arrow) Image taken at 7 days post inoculation (Colored with DAPI).

Fig. 14

The glioblastoma induced tumor at 28 days post–inoculation (14 A). The microscopic aspect (14 B). Colored with hematoxilin–eozina.