Immortalization of human normal and NF1 neurofibroma Schwann cells.
Neurofibromas, which are benign Schwann cell tumors, are the hallmark feature in the autosomal dominant condition neurofibromatosis 1 (NF1) and are associated with biallelic loss of NF1 gene function. There is a need for effective therapies for neurofibromas, particularly the larger, plexiform neurofibromas. Tissue culture is an important tool for research. However, it is difficult to derive enriched human Schwann cell cultures, and most enter replicative senescence after 6-10 passages, impeding cell-based research in NF1. Through exogenous expression of human telomerase reverse transcriptase and murine cyclin-dependent kinase (mCdk4), normal (NF1 wild-type), neurofibroma-derived Schwann cells heterozygous for NF1 mutation, and neurofibroma-derived Schwann cells homozygous for NF1 mutation were immortalized, including some matched samples from the same NF1 patient. Initial experiments employed retroviral vectors, while subsequent work utilized lentiviral vectors carrying these genes because of improved efficiency. Expression of both transgenes was required for immortalization. Molecular and immunohistochemical analysis indicated that these cell lines are of Schwann cell lineage and have a range of phenotypes, many of which are consistent with their primary cultures. This is the first report of immortalization and detailed characterization of multiple human NF1 normal nerve and neurofibroma-derived Schwann cell lines, which will be highly useful research tools to study NF1 and other Schwann tumor biology and conditions.
Department of Molecular Genetics and Microbiology, University of Florida College of Medicine, Gainesville, FL, USA.
Hua
H
Li
Department of Molecular Genetics and Microbiology, University of Florida College of Medicine, Gainesville, FL, USA.
Lung-Ji
LJ
Chang
Department of Pediatrics, Child Health Research Institute, University of Florida College of Medicine, Gainesville, FL, USA.
Debbie R
DR
Neubauer
Department of Pediatrics, Child Health Research Institute, University of Florida College of Medicine, Gainesville, FL, USA.|Department of Neuroscience, University of Florida College of Medicine, Gainesville, FL, USA.|University of Florida Health Cancer Center, University of Florida, Gainesville, FL, USA.
David F
DF
Muir
Department of Molecular Genetics and Microbiology, University of Florida College of Medicine, Gainesville, FL, USA.|University of Florida Health Cancer Center, University of Florida, Gainesville, FL, USA.|University of Florida Genetics Institute, University of Florida, Gainesville, FL, USA.
Margaret R
MR
Wallace
D000818:Animals; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D051358:Cyclin-Dependent Kinase 4 / Q000235:genetics; D006801:Humans; D051379:Mice; D009368:Neoplasm Transplantation; D009456:Neurofibromatosis 1; D012583:Schwann Cells; D019098:Telomerase / Q000235:genetics
C495902:Cdk4 protein, mouse; D051358:Cyclin-Dependent Kinase 4; D019098:Telomerase