Figure 2.

Morphological, genetic, and functional traits of tumor subclones are preserved in vitro and in vivo. (A) H&E stained original tissue biopsy (case GNV019) diagnosed as glioblastoma. Typical necrosis, microvascular proliferation, and mitotically active glial cells are present. Note the rare mGCs (example highlighted in box). (B) Phase contrast appearance of GNV019 parental cells, and (C) respective growth kinetics for 35 passages in vitro. (D) Scatterplot of log2-transformed whole genome gene expression data from passage 5 and passage 10 GNV019 parental cells revealing high correlation. 96.8% of all expressed genes are found within the 2-fold demarcated area. (E) LogR ratio plots illustrating glioblastoma-typic aberrations (28, 29) from passage 3 vs. passage 10 GNV019 parental cells. Note the overlap. (F) Serial coronal T2-weighted MRI of an ex vivo recipient SCID-beige mouse whole brain demonstrating development of a large T2-intermediate mass infiltrating left frontal cortex and subjacent basal ganglia 44 days after xenografting GNV019 parental cells (asterisk indicates original transplant site). There is herniation and midline shift from substantial mass effect (G) Microscopic appearance of the respective xenograft (H&E, tissue section). Note the similarity of pathology to the original patient specimen (Fig. 2A). One of the rare mGCs is exposed (box). (H) Subclones derived from GNV019 parental cells at passage 5. Upper panel depicts their phase contrast appearance in vitro. Lower panel: H&E stains expose distinct xenograft morphologies. CL2 cells were not tumorigenic. (I) Distribution of mGCs in respective xenografts. For quantification, a mean of 10,145 cells were counted in 3–10 random 40× fields per case. PA=GNV019 parental cells. Scale bars: A, 250 µm; B, H 50 µm; G, 100 µm. See also Supplementary Fig. S2.