title: Generation of an immortalized astrocyte cell line from H-2Kb-tsA58 mice to study the role of astrocytes in brain metastasis.
abstract: Astrocytes play a critical role in maintaining cerebral homeostasis and their dysregulation is thought to contribute to the pathogenesis of several diseases, including brain cancer and metastasis. Similar to the human disease, we found that lung and melanoma metastases in the mouse brain are accompanied by a reactive gliosis. To begin to study the biology of astrocytes and examine how these cells might contribute to metastasis formation and progression in the brain, we generated a conditionally immortal astrocyte cell line from H-2Kb-tsA58 mice. Astrocytes grown in culture expressed glial fibrillary acid protein (GFAP), glutamate receptor 1, and the N-methyl-D-aspartate (NMDA) receptor. Astrocytes also expressed the glial-specific transporters excitatory amino acid transporter 1 (EAAT1) and EAAT2. Astrocytes grown under permissive conditions (33 degrees C) expressed SV40 large T antigen and had a doubling time of 36 h, whereas expression of SV40 large T antigen was negligible in astrocytes grown at 37 degrees C for 72 h, which coincided with a plateau in cell division. In a co-culture assay with human lung adenocarcinoma cells (PC14-PE6), astrocytes activated programs in the tumor cells that signal for cell division and survival. Hence, the immortalized cell line will be useful for studying the role of astrocytes in disease processes in the brain, such as metastasis.
authors.affiliation: Department of Cancer Biology, Unit 173, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA. rlangley@mdanderson.org
authors.forename: Robert R
authors.initials: RR
authors.lastname: Langley
authors.forename: Dominic
authors.initials: D
authors.lastname: Fan
authors.forename: Lixia
authors.initials: L
authors.lastname: Guo
authors.forename: Chenyu
authors.initials: C
authors.lastname: Zhang
authors.forename: Qingtang
authors.initials: Q
authors.lastname: Lin
authors.forename: Emily C
authors.initials: EC
authors.lastname: Brantley
authors.forename: Joseph H
authors.initials: JH
authors.lastname: McCarty
authors.forename: Isaiah J
authors.initials: IJ
authors.lastname: Fidler
mesh_terms: D000230:Adenocarcinoma / Q000473:pathology; D000818:Animals; D000831:Animals, Newborn; D015703:Antigens, CD / Q000378:metabolism; D015214:Antigens, Differentiation, Myelomonocytic / Q000378:metabolism; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics*; D001253:Astrocytes / Q000378:metabolism / Q000473:pathology*; D001932:Brain Neoplasms / Q000378:metabolism / Q000556:secondary*; D002450:Cell Communication; D002460:Cell Line; D049109:Cell Proliferation; D002470:Cell Survival; D002472:Cell Transformation, Viral / Q000235:genetics*; D018920:Coculture Techniques; D027341:Excitatory Amino Acid Transporter 1 / Q000378:metabolism; D027342:Excitatory Amino Acid Transporter 2 / Q000378:metabolism; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D006183:H-2 Antigens / Q000235:genetics*; D006801:Humans; D008175:Lung Neoplasms / Q000473:pathology; D008545:Melanoma / Q000473:pathology; D051379:Mice; D008809:Mice, Inbred C3H; D008819:Mice, Nude; D008822:Mice, Transgenic; D016194:Receptors, N-Methyl-D-Aspartate / Q000378:metabolism; D013696:Temperature; D013997:Time Factors; D000097382:CD68 Molecule
chemical_list: D015703:Antigens, CD; D015214:Antigens, Differentiation, Myelomonocytic; D000952:Antigens, Polyomavirus Transforming; D027341:Excitatory Amino Acid Transporter 1; D027342:Excitatory Amino Acid Transporter 2; D005904:Glial Fibrillary Acidic Protein; D006183:H-2 Antigens; D016194:Receptors, N-Methyl-D-Aspartate; C067980:CD68 antigen, human; D000097382:CD68 Molecule; C081026:CD68 protein, mouse; C032876:H-2Kb protein, mouse; C493396:Slc1a2 protein, mouse; C493393:Slc1a3 protein, mouse