Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2012 Apr;107(2):269-80.
doi: 10.1007/s11060-011-0756-5. Epub 2011 Nov 27.

Novel Cell Lines Established From Pediatric Brain Tumors

Affiliations
Free PMC article

Novel Cell Lines Established From Pediatric Brain Tumors

Jingying Xu et al. J Neurooncol. .
Free PMC article

Abstract

The paucity of cell culture models for childhood brain tumors prompted us to establish pediatric cell lines for use in biological experiments and preclinical developmental therapeutic studies. Three cell lines were established, CHLA-200 (GBM), CHLA-259 (anaplastic medulloblastoma) and CHLA-266 (atypical teratoid rhabdoid tumor, AT/RT). Consistent with an AT/RT origin, CHLA-266 lacked INI1 expression and had monosomy 22. All lines had unique DNA short tandem repeat "fingerprints" matching that of the patient's tumor tissue and were adherent on tissue culture plastic, but differed in morphology and doubling times. CHLA-200 had a silent mutation in TP53. CHLA-259 and CHLA-266 had wild-type TP53. All three lines were relatively resistant to multiple drugs when compared to the DAOY medulloblastoma cell line, using the DIMSCAN fluorescence digital image microscopy cytotoxicity assay. RNA expression of MYC and MYCN were quantified using RT-PCR (Taqman). CHLA-200 expressed MYC, DAOY and CHLA-259 expressed MYCN, and CHLA-266 expressed both MYCN and MYC. CHLA-200 was only tumorigenic subcutaneously, but CHLA-259 and CHLA-266 were tumorigenic both subcutaneously and in brains of NOD/SCID mice. Immunohistochemistry of the xenografts revealed GFAP staining in CHLA-200 and PGP 9.5 staining in CHLA-259 and CHLA-266 tumors. As expected, INI1 expression was lacking in CHLA-266 (AT/RT). These three new cell lines will provide useful models for research of pediatric brain tumors.

Figures

Fig. 1
Fig. 1
Morphology of pediatric brain tumor cell lines in log phase growth. Doubling time (DT) of each cell line is shown in the panel
Fig. 2
Fig. 2
CHLA-200, CHLA-259 and CHLA-266 show higher resistance to most chemotherapy drugs tested as compared to DAOY cells. Cytotoxicity was analyzed after 4 days exposure using the DIM-SCAN assay to cisplatin (CDDP), cyclophosphamide (as 4-hydroperoxycyclophosphamide, 4-HC), etoposide (ETOP), melphalan (L-PAM), topotecan (TPT), vincristin (VINC), and ferentinide (4-HPR). For each drug concentration n = 12
Fig. 3
Fig. 3
Melphalan (L-PAM) induced variable p53 expression and signaling in CHLA-200, CHLA-259, CHLA-266, and DAOY cell lines. Cells (1 × 106) plated in six well plates were incubated with L-PAM (6 µg/ml) or vehicle in 2 ml medium. After 16 h cells were collected, cell lysates (20 µg for p53 and MDM2 and 10 µg for p21) were resolved by SDS-PAGE and detected by western blotting. Fold increase in expression in presence of drug compared to control was calculated by densitometry using actin as loading control, and is denoted below the blots. This is a representative experiment of three with similar results
Fig. 4
Fig. 4
The new pediatric brain tumor cell lines express telomerase and MYC genes. Quantitative RT-PCR of the three cell lines and DAOY: a mRNA expression of MYC and MYCN oncogenes, b mRNA of telomerase: hTR and hTERT. Bars represent means ± SD of triplicate samples in one representative experiment out of three experiments with similar results
Fig. 5
Fig. 5
CHLA-259 and CHLA-266 form intracranial tumors in mice. a MRI images of tumors in mouse brains: Left panel CHLA-266, day 82, 2.1 × 2.0 mm tumor, Right panel CHLA-259, day 120, 1.9 × 2.4 mm tumor. b H&E stains of the original patient tumors (top panels) and the mouse brain tumors from orthotopic implantation of their respective cell lines (bottom). Original magnification is 4009. Arrows Apoptotic cells in CHLA-259, Arrowheads typical rhabdoid cells in CHLA-266. c Immunohistochemical staining for synaptophysin, PGP 9.5, GFAP, vimentin and INI1 of the mouse xenograft of CHLA-200 (subcutaneous), CHLA-266 and CHLA-259 (both intracranial) compared to normal brain

Similar articles

See all similar articles

Cited by 22 articles

See all "Cited by" articles

Publication types

MeSH terms

Feedback