Table 1.
Clinical data of the patients.
Fig 1.
Summary of tissue processing and cell line establishment protocol.
Table 2.
Cell line establishment criteria.
Fig 2.
Outcome of short tandem repeat DNA profiling of the established cell lines showed the unique profile of each line.
Twenty markers were assayed and an overlap of 16 or more markers (80%) was considered an indication of cross-contamination. HeLa and K562 were included both as quality controls and to show lack of contamination from these ubiquitous cell lines.
Fig 3.
Cell line growth and multipotency.
(A-B) Phase contrast images of oncospheres from JHU-0879 (A) and JHU-0937 (B). (C-F) Immunofluorescence images demonstrating the multipotency of the oncospheres. (C-D) Astrocytic-like cells stained for glial fibrillary acidic protein in lines JHU-0879 (C) and JHU-0937 (D). (E-F) Neuronal-like cells stained for class III Ī²-tubulin in lines JHU-0879 (E) and JHU-0937 (F). All images taken at 400x.
Fig 4.
H&E stains of orthotopic tumors formed in athymic mice, showing hallmark characteristics of GBMs.
(A and B) All of the lines showed diffuse invasion of normal parenchymal. The hallmark features of glioblastoma were identified in the brains of mice injected with oncosphere lines, including increased mitotic activity (C, arrows), necrosis (D, designated āNā), and vascular proliferation (E, arrows). The oncosphere lines also demonstrated other histologic features specific to invasive gliomas, including spread through white matter tracts (F), neuronal satellitosis (G), and subventricular tumor formation (H). Original magnification for panel A was 25x; for F and H, 50x; for B and D, 100x; and for C, E, and G, 200x.
Fig 5.
H&E stains of primary tumor tissue and orthotopic tumor from the GBM variant cell lines showing specific features of each variant.
The JHH-505 primary tumor has features of a GBM such as vascular proliferation and dense cellular areas (A) while certain areas contain round, regular oligodendroglial appearing cells (B). The JHH-505 xenograft tumor also contains histological characteristics consistent with GBM-O, including dense (C) and less cellular (D) regions with oligodendroglial appearing cells. The JHU-0879 primary tumor has areas consistent with PNET with hypercellularity containing cells with scant cytoplasm and prominent nucleoli (E). The JHU-0879 xenograft also has PNET features, such as densely packed cells and prominent nucleoli (F). Arrows highlight mitotic or apoptotic cells. Magnification for panel A is 100X, panel B-F are 400X.