title: Glioblastoma cell line-derived spheres in serum‑containing medium versus serum-free medium: a comparison of cancer stem cell properties.
abstract: In addition to the primary culturing of cancer stem cells (CSCs) from tumor             tissues, CSCs are found in established tumor cell lines. However, it is unclear             how culture conditions affect CSC enrichment. Additionally, the differentiation             potential of cell line-derived CSCs has not been well studied. In our study, the             glioblastoma cell lines LN229, T98G, U251n and U87, were cultured as spheres in             serum-containing medium (serum spheres) or serum-free medium (serum-free spheres).             We found that LN229 and U251n cells expressed multiple stem cell markers such             as Nestin, Sox2, Musashi-1 and CD44, and their serum spheres expressed even higher             levels of Nestin, Sox2 and Musashi-1 compared to monolayer cells and serum‑free             spheres. LN229 and U251n cells showed higher migration and colony formation potential             compared to T98G and U87 cells, which did not express Nestin, Sox2 and Musashi-1.             Serum spheres of LN229 and U251n cells also exhibited higher resistance to temozolomide             compared to serum-free spheres. All tumor cell lines showed neuronal differentiation (Tuj-1             positive). Only U251n serum spheres exhibited both astrocytic (GFAP‑positive)             and neuronal differentiation. We conclude that sphere culture in serum-containing             medium provides the most efficient enrichment of cancer stem cells. U251n cells             are distinguished from other tumor cells due to their potential for multilineage             differentiation.
authors.affiliation: Department of Neurosurgery, Henry Ford Health System, Detroit,                     MI 48202, USA.
authors.forename: Xin
authors.initials: X
authors.lastname: Hong
authors.forename: Khalil
authors.initials: K
authors.lastname: Chedid
authors.forename: Steven N
authors.initials: SN
authors.lastname: Kalkanis
mesh_terms: D002454:Cell Differentiation / Q000187:drug effects; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation / Q000187:drug effects; D016895:Culture Media, Serum-Free; D003606:Dacarbazine / Q000031:analogs & derivatives / Q000494:pharmacology / Q000633:toxicity; D005909:Glioblastoma / Q000378:metabolism* / Q000473:pathology*; D006801:Humans; D007381:Intermediate Filament Proteins / Q000096:biosynthesis; D014411:Neoplastic Stem Cells / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology*; D009419:Nerve Tissue Proteins / Q000096:biosynthesis; D064231:Nestin; D016601:RNA-Binding Proteins / Q000096:biosynthesis; D055748:SOXB1 Transcription Factors / Q000096:biosynthesis; D018874:Spheroids, Cellular; D000077204:Temozolomide; D014407:Tumor Cells, Cultured; D014410:Tumor Stem Cell Assay
chemical_list: D016895:Culture Media, Serum-Free; D007381:Intermediate Filament Proteins; C491297:MSI1 protein, human; C577484:NES protein, human; D009419:Nerve Tissue Proteins; D064231:Nestin; D016601:RNA-Binding Proteins; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; D003606:Dacarbazine; D000077204:Temozolomide