Neuro-Oncology: Novel cell lines established from pediatric brain tumors

Wednesday, November 30, 2011

Novel cell lines established from pediatric brain tumors

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.

Content Type Journal Article
Category Laboratory Investigation - Human/Animal Tissue
Pages 1-12
DOI 10.1007/s11060-011-0756-5
Authors
- Jingying Xu, Developmental Therapeutics Program, Division of Hematology-Oncology, USC-CHLA Institute for Pediatric Clinical Research, Children's Hospital Los Angeles, Los Angeles, CA 90027, USA
- Anat Erdreich-Epstein, Department of Pediatrics and Pathology, Keck School of Medicine, University of Southern California, Los Angeles, CA 90027, USA
- Ignacio Gonzalez-Gomez, Children's Hospital Los Angeles, Los Angeles, CA 90027, USA
- Elizabeth Y. Melendez, Children's Hospital Los Angeles, Los Angeles, CA 90027, USA
- Goar Smbatyan, Children's Hospital Los Angeles, Los Angeles, CA 90027, USA
- Rex A. Moats, Children's Hospital Los Angeles, Los Angeles, CA 90027, USA
- Michael Rosol, Children's Hospital Los Angeles, Los Angeles, CA 90027, USA
- Jaclyn A. Biegel, Division of Human Genetics, The Children's Hospital of Philadelphia, Philadelphia, PA 19104, USA
- C. Patrick Reynolds, Developmental Therapeutics Program, Division of Hematology-Oncology, USC-CHLA Institute for Pediatric Clinical Research, Children's Hospital Los Angeles, Los Angeles, CA 90027, USA