Culture conditions tailored to the cell of origin are critical for maintaining native properties and tumorigenicity of glioma cells.
Cell culture plays a pivotal role in cancer research. However, culture-induced changes in biological properties of tumor cells profoundly affect research reproducibility and translational potential. Establishing culture conditions tailored to the cancer cell of origin could resolve this problem. For glioma research, it has been previously shown that replacing serum with defined growth factors for neural stem cells (NSCs) greatly improved the retention of gene expression profile and tumorigenicity. However, among all molecular subtypes of glioma, our laboratory and others have previously shown that the oligodendrocyte precursor cell (OPC) rather than the NSC serves as the cell of origin for the proneural subtype, raising questions regarding the suitability of NSC-tailored media for culturing proneural glioma cells.



OPC-originated mouse glioma cells were cultured in conditions for normal OPCs or NSCs, respectively, for multiple passages. Gene expression profiles, morphologies, tumorigenicity, and drug responsiveness of cultured cells were examined in comparison with freshly isolated tumor cells.



OPC media-cultured glioma cells maintained tumorigenicity, gene expression profiles, and morphologies similar to freshly isolated tumor cells. In contrast, NSC-media cultured glioma cells gradually lost their OPC features and most tumor-initiating ability and acquired heightened sensitivity to temozolomide.



To improve experimental reproducibility and translational potential of glioma research, it is important to identify the cell of origin, and subsequently apply this knowledge to establish culture conditions that allow the retention of native properties of tumor cells.
Department of Microbiology, Immunology, and Cancer Biology, University of Virginia, Charlottesville, Virginia (P.F.L., C.L., A.R.H., H.Z.); Department of Neurological Surgery, University of Virginia, Charlottesville, Virginia (M.E.S., A.A., D.S.); Division of Neuropathology, Department of Pathology, Charlottesville, Virginia (M.B.S.L., J.W.M.); Department of Neurology, School of Medicine, University of Virginia, Charlottesville, Virginia (D.S.); Department of Pathology and Pathological Physiology, Center for Cancer Research, Zhejiang University School of Medicine, Zhejiang Diseases Proteomics Key Laboratory, Hangzhou, China (C.L.); Department of Biophysics and Center of Biotechnology, Federal University of Rio Grande do Sul, Porto Alegre, Brazil (P.F.L., D.C.M., G.L.); Department of Neurosurgery, Changzheng Hospital, Second Affiliated Hospital of Second Military Medical University, Shanghai, China (H.H., Y.-C.L.); Department of Pathology, Xiang-ya Hospital, Central South University, Changsha, Hunan, China (H.-Y.Z.).
Pítia F
PF
Ledur
Department of Microbiology, Immunology, and Cancer Biology, University of Virginia, Charlottesville, Virginia (P.F.L., C.L., A.R.H., H.Z.); Department of Neurological Surgery, University of Virginia, Charlottesville, Virginia (M.E.S., A.A., D.S.); Division of Neuropathology, Department of Pathology, Charlottesville, Virginia (M.B.S.L., J.W.M.); Department of Neurology, School of Medicine, University of Virginia, Charlottesville, Virginia (D.S.); Department of Pathology and Pathological Physiology, Center for Cancer Research, Zhejiang University School of Medicine, Zhejiang Diseases Proteomics Key Laboratory, Hangzhou, China (C.L.); Department of Biophysics and Center of Biotechnology, Federal University of Rio Grande do Sul, Porto Alegre, Brazil (P.F.L., D.C.M., G.L.); Department of Neurosurgery, Changzheng Hospital, Second Affiliated Hospital of Second Military Medical University, Shanghai, China (H.H., Y.-C.L.); Department of Pathology, Xiang-ya Hospital, Central South University, Changsha, Hunan, China (H.-Y.Z.) chongliu77@zju.edu.cn hz9s@virginia.edu.
Chong
C
Liu
Department of Microbiology, Immunology, and Cancer Biology, University of Virginia, Charlottesville, Virginia (P.F.L., C.L., A.R.H., H.Z.); Department of Neurological Surgery, University of Virginia, Charlottesville, Virginia (M.E.S., A.A., D.S.); Division of Neuropathology, Department of Pathology, Charlottesville, Virginia (M.B.S.L., J.W.M.); Department of Neurology, School of Medicine, University of Virginia, Charlottesville, Virginia (D.S.); Department of Pathology and Pathological Physiology, Center for Cancer Research, Zhejiang University School of Medicine, Zhejiang Diseases Proteomics Key Laboratory, Hangzhou, China (C.L.); Department of Biophysics and Center of Biotechnology, Federal University of Rio Grande do Sul, Porto Alegre, Brazil (P.F.L., D.C.M., G.L.); Department of Neurosurgery, Changzheng Hospital, Second Affiliated Hospital of Second Military Medical University, Shanghai, China (H.H., Y.-C.L.); Department of Pathology, Xiang-ya Hospital, Central South University, Changsha, Hunan, China (H.-Y.Z.).
Hua
H
He
Department of Microbiology, Immunology, and Cancer Biology, University of Virginia, Charlottesville, Virginia (P.F.L., C.L., A.R.H., H.Z.); Department of Neurological Surgery, University of Virginia, Charlottesville, Virginia (M.E.S., A.A., D.S.); Division of Neuropathology, Department of Pathology, Charlottesville, Virginia (M.B.S.L., J.W.M.); Department of Neurology, School of Medicine, University of Virginia, Charlottesville, Virginia (D.S.); Department of Pathology and Pathological Physiology, Center for Cancer Research, Zhejiang University School of Medicine, Zhejiang Diseases Proteomics Key Laboratory, Hangzhou, China (C.L.); Department of Biophysics and Center of Biotechnology, Federal University of Rio Grande do Sul, Porto Alegre, Brazil (P.F.L., D.C.M., G.L.); Department of Neurosurgery, Changzheng Hospital, Second Affiliated Hospital of Second Military Medical University, Shanghai, China (H.H., Y.-C.L.); Department of Pathology, Xiang-ya Hospital, Central South University, Changsha, Hunan, China (H.-Y.Z.).
Alexandra R
AR
Harris
Department of Microbiology, Immunology, and Cancer Biology, University of Virginia, Charlottesville, Virginia (P.F.L., C.L., A.R.H., H.Z.); Department of Neurological Surgery, University of Virginia, Charlottesville, Virginia (M.E.S., A.A., D.S.); Division of Neuropathology, Department of Pathology, Charlottesville, Virginia (M.B.S.L., J.W.M.); Department of Neurology, School of Medicine, University of Virginia, Charlottesville, Virginia (D.S.); Department of Pathology and Pathological Physiology, Center for Cancer Research, Zhejiang University School of Medicine, Zhejiang Diseases Proteomics Key Laboratory, Hangzhou, China (C.L.); Department of Biophysics and Center of Biotechnology, Federal University of Rio Grande do Sul, Porto Alegre, Brazil (P.F.L., D.C.M., G.L.); Department of Neurosurgery, Changzheng Hospital, Second Affiliated Hospital of Second Military Medical University, Shanghai, China (H.H., Y.-C.L.); Department of Pathology, Xiang-ya Hospital, Central South University, Changsha, Hunan, China (H.-Y.Z.).
Darlan C
DC
Minussi
Department of Microbiology, Immunology, and Cancer Biology, University of Virginia, Charlottesville, Virginia (P.F.L., C.L., A.R.H., H.Z.); Department of Neurological Surgery, University of Virginia, Charlottesville, Virginia (M.E.S., A.A., D.S.); Division of Neuropathology, Department of Pathology, Charlottesville, Virginia (M.B.S.L., J.W.M.); Department of Neurology, School of Medicine, University of Virginia, Charlottesville, Virginia (D.S.); Department of Pathology and Pathological Physiology, Center for Cancer Research, Zhejiang University School of Medicine, Zhejiang Diseases Proteomics Key Laboratory, Hangzhou, China (C.L.); Department of Biophysics and Center of Biotechnology, Federal University of Rio Grande do Sul, Porto Alegre, Brazil (P.F.L., D.C.M., G.L.); Department of Neurosurgery, Changzheng Hospital, Second Affiliated Hospital of Second Military Medical University, Shanghai, China (H.H., Y.-C.L.); Department of Pathology, Xiang-ya Hospital, Central South University, Changsha, Hunan, China (H.-Y.Z.).
Hai-Yan
HY
Zhou
Department of Microbiology, Immunology, and Cancer Biology, University of Virginia, Charlottesville, Virginia (P.F.L., C.L., A.R.H., H.Z.); Department of Neurological Surgery, University of Virginia, Charlottesville, Virginia (M.E.S., A.A., D.S.); Division of Neuropathology, Department of Pathology, Charlottesville, Virginia (M.B.S.L., J.W.M.); Department of Neurology, School of Medicine, University of Virginia, Charlottesville, Virginia (D.S.); Department of Pathology and Pathological Physiology, Center for Cancer Research, Zhejiang University School of Medicine, Zhejiang Diseases Proteomics Key Laboratory, Hangzhou, China (C.L.); Department of Biophysics and Center of Biotechnology, Federal University of Rio Grande do Sul, Porto Alegre, Brazil (P.F.L., D.C.M., G.L.); Department of Neurosurgery, Changzheng Hospital, Second Affiliated Hospital of Second Military Medical University, Shanghai, China (H.H., Y.-C.L.); Department of Pathology, Xiang-ya Hospital, Central South University, Changsha, Hunan, China (H.-Y.Z.).
Mark E
ME
Shaffrey
Department of Microbiology, Immunology, and Cancer Biology, University of Virginia, Charlottesville, Virginia (P.F.L., C.L., A.R.H., H.Z.); Department of Neurological Surgery, University of Virginia, Charlottesville, Virginia (M.E.S., A.A., D.S.); Division of Neuropathology, Department of Pathology, Charlottesville, Virginia (M.B.S.L., J.W.M.); Department of Neurology, School of Medicine, University of Virginia, Charlottesville, Virginia (D.S.); Department of Pathology and Pathological Physiology, Center for Cancer Research, Zhejiang University School of Medicine, Zhejiang Diseases Proteomics Key Laboratory, Hangzhou, China (C.L.); Department of Biophysics and Center of Biotechnology, Federal University of Rio Grande do Sul, Porto Alegre, Brazil (P.F.L., D.C.M., G.L.); Department of Neurosurgery, Changzheng Hospital, Second Affiliated Hospital of Second Military Medical University, Shanghai, China (H.H., Y.-C.L.); Department of Pathology, Xiang-ya Hospital, Central South University, Changsha, Hunan, China (H.-Y.Z.).
Ashok
A
Asthagiri
Department of Microbiology, Immunology, and Cancer Biology, University of Virginia, Charlottesville, Virginia (P.F.L., C.L., A.R.H., H.Z.); Department of Neurological Surgery, University of Virginia, Charlottesville, Virginia (M.E.S., A.A., D.S.); Division of Neuropathology, Department of Pathology, Charlottesville, Virginia (M.B.S.L., J.W.M.); Department of Neurology, School of Medicine, University of Virginia, Charlottesville, Virginia (D.S.); Department of Pathology and Pathological Physiology, Center for Cancer Research, Zhejiang University School of Medicine, Zhejiang Diseases Proteomics Key Laboratory, Hangzhou, China (C.L.); Department of Biophysics and Center of Biotechnology, Federal University of Rio Grande do Sul, Porto Alegre, Brazil (P.F.L., D.C.M., G.L.); Department of Neurosurgery, Changzheng Hospital, Second Affiliated Hospital of Second Military Medical University, Shanghai, China (H.H., Y.-C.L.); Department of Pathology, Xiang-ya Hospital, Central South University, Changsha, Hunan, China (H.-Y.Z.).
Maria Beatriz S
MB
Lopes
Department of Microbiology, Immunology, and Cancer Biology, University of Virginia, Charlottesville, Virginia (P.F.L., C.L., A.R.H., H.Z.); Department of Neurological Surgery, University of Virginia, Charlottesville, Virginia (M.E.S., A.A., D.S.); Division of Neuropathology, Department of Pathology, Charlottesville, Virginia (M.B.S.L., J.W.M.); Department of Neurology, School of Medicine, University of Virginia, Charlottesville, Virginia (D.S.); Department of Pathology and Pathological Physiology, Center for Cancer Research, Zhejiang University School of Medicine, Zhejiang Diseases Proteomics Key Laboratory, Hangzhou, China (C.L.); Department of Biophysics and Center of Biotechnology, Federal University of Rio Grande do Sul, Porto Alegre, Brazil (P.F.L., D.C.M., G.L.); Department of Neurosurgery, Changzheng Hospital, Second Affiliated Hospital of Second Military Medical University, Shanghai, China (H.H., Y.-C.L.); Department of Pathology, Xiang-ya Hospital, Central South University, Changsha, Hunan, China (H.-Y.Z.).
David
D
Schiff
Department of Microbiology, Immunology, and Cancer Biology, University of Virginia, Charlottesville, Virginia (P.F.L., C.L., A.R.H., H.Z.); Department of Neurological Surgery, University of Virginia, Charlottesville, Virginia (M.E.S., A.A., D.S.); Division of Neuropathology, Department of Pathology, Charlottesville, Virginia (M.B.S.L., J.W.M.); Department of Neurology, School of Medicine, University of Virginia, Charlottesville, Virginia (D.S.); Department of Pathology and Pathological Physiology, Center for Cancer Research, Zhejiang University School of Medicine, Zhejiang Diseases Proteomics Key Laboratory, Hangzhou, China (C.L.); Department of Biophysics and Center of Biotechnology, Federal University of Rio Grande do Sul, Porto Alegre, Brazil (P.F.L., D.C.M., G.L.); Department of Neurosurgery, Changzheng Hospital, Second Affiliated Hospital of Second Military Medical University, Shanghai, China (H.H., Y.-C.L.); Department of Pathology, Xiang-ya Hospital, Central South University, Changsha, Hunan, China (H.-Y.Z.).
Yi-Cheng
YC
Lu
Department of Microbiology, Immunology, and Cancer Biology, University of Virginia, Charlottesville, Virginia (P.F.L., C.L., A.R.H., H.Z.); Department of Neurological Surgery, University of Virginia, Charlottesville, Virginia (M.E.S., A.A., D.S.); Division of Neuropathology, Department of Pathology, Charlottesville, Virginia (M.B.S.L., J.W.M.); Department of Neurology, School of Medicine, University of Virginia, Charlottesville, Virginia (D.S.); Department of Pathology and Pathological Physiology, Center for Cancer Research, Zhejiang University School of Medicine, Zhejiang Diseases Proteomics Key Laboratory, Hangzhou, China (C.L.); Department of Biophysics and Center of Biotechnology, Federal University of Rio Grande do Sul, Porto Alegre, Brazil (P.F.L., D.C.M., G.L.); Department of Neurosurgery, Changzheng Hospital, Second Affiliated Hospital of Second Military Medical University, Shanghai, China (H.H., Y.-C.L.); Department of Pathology, Xiang-ya Hospital, Central South University, Changsha, Hunan, China (H.-Y.Z.).
James W
JW
Mandell
Department of Microbiology, Immunology, and Cancer Biology, University of Virginia, Charlottesville, Virginia (P.F.L., C.L., A.R.H., H.Z.); Department of Neurological Surgery, University of Virginia, Charlottesville, Virginia (M.E.S., A.A., D.S.); Division of Neuropathology, Department of Pathology, Charlottesville, Virginia (M.B.S.L., J.W.M.); Department of Neurology, School of Medicine, University of Virginia, Charlottesville, Virginia (D.S.); Department of Pathology and Pathological Physiology, Center for Cancer Research, Zhejiang University School of Medicine, Zhejiang Diseases Proteomics Key Laboratory, Hangzhou, China (C.L.); Department of Biophysics and Center of Biotechnology, Federal University of Rio Grande do Sul, Porto Alegre, Brazil (P.F.L., D.C.M., G.L.); Department of Neurosurgery, Changzheng Hospital, Second Affiliated Hospital of Second Military Medical University, Shanghai, China (H.H., Y.-C.L.); Department of Pathology, Xiang-ya Hospital, Central South University, Changsha, Hunan, China (H.-Y.Z.).
Guido
G
Lenz
Department of Microbiology, Immunology, and Cancer Biology, University of Virginia, Charlottesville, Virginia (P.F.L., C.L., A.R.H., H.Z.); Department of Neurological Surgery, University of Virginia, Charlottesville, Virginia (M.E.S., A.A., D.S.); Division of Neuropathology, Department of Pathology, Charlottesville, Virginia (M.B.S.L., J.W.M.); Department of Neurology, School of Medicine, University of Virginia, Charlottesville, Virginia (D.S.); Department of Pathology and Pathological Physiology, Center for Cancer Research, Zhejiang University School of Medicine, Zhejiang Diseases Proteomics Key Laboratory, Hangzhou, China (C.L.); Department of Biophysics and Center of Biotechnology, Federal University of Rio Grande do Sul, Porto Alegre, Brazil (P.F.L., D.C.M., G.L.); Department of Neurosurgery, Changzheng Hospital, Second Affiliated Hospital of Second Military Medical University, Shanghai, China (H.H., Y.-C.L.); Department of Pathology, Xiang-ya Hospital, Central South University, Changsha, Hunan, China (H.-Y.Z.).
Hui
H
Zong
D000818:Animals; D015153:Blotting, Western; D018929:Cell Culture Techniques / Q000379:methods*; D016000:Cluster Analysis; D005455:Fluorescent Antibody Technique; D005910:Glioma / Q000473:pathology*; D064593:Heterografts; D006801:Humans; D051379:Mice; D014411:Neoplastic Stem Cells / Q000473:pathology*; D058953:Neural Stem Cells / Q000473:pathology*; D009474:Neurons / Q000473:pathology; D009836:Oligodendroglia / Q000473:pathology*; D060888:Real-Time Polymerase Chain Reaction; D046888:Tissue Array Analysis; D059467:Transcriptome; D014407:Tumor Cells, Cultured
cell culture; glioma; neural stem cell (NSC); oligodendrocyte precursor cell (OPC); research reproducibility