Long-term maintenance of brain tumor stem cell properties under at non-adherent and adherent culture conditions

Inagaki, Akihito; Soeda, Akio; Oka, Naoki; Kitajima, Hideomi; Nakagawa, Jiro; Motohashi, Tsutomu; Kunisada, Takahiro; Iwama, Toru

doi:10.1016/j.bbrc.2007.07.037

Cited by 45 publications

(61 citation statements)

References 18 publications

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“…pivotal role in tumor initiation, growth, chemoresistance and recurrence (34,35). to establish a more detailed understanding, functional assays of this subpopulation have been evaluated, and consequently long-term cultures of cancer stem-like cells have been established recently in several types of malignancies (9,32,(36)(37)(38).…”

Section: Discussionmentioning

confidence: 99%

Maintenance of retinal cancer stem cell-like properties through long-term serum-free culture from human retinoblastoma

Jiao

2011

Oncol Rep

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Abstract. previous studies have demonstrated that a small population of cancer stem cell-like cells exists in retinoblastoma. to provide a model for studying this population, we sought to establish a long-term culture from human retinoblastoma that have cancer stem cell-like properties. Fresh tumor tissue was digested and cultured in serum-free medium. tumor spheres formed and were passaged continuously. stem cell properties were examined through immunostaining, real-time quantitative rt-pCr and chemoresistance assay. tumorigenicity of the tumor sphere-forming cells was confirmed by xenograft experiments. Furthermore, we examined the expression of cell surface markers CD44 and CD133. tumor cells expanded as floating spheres for more than 30 passages. Sphere-forming cells overexpressed stem cell genes Oct-4, Nestin and Pax6. Immunostaining of spheres showed positivity for Nestin, pax6 and also ABCG2. In contrast, differentiated cells derived from these spheres expressed high levels of mature retinal cell markers MAp2, GFAp, recoverin, Opsin B and Nrl, and showed immunoreactivity for NF200, GFAp, recoverin and pKCα. Furthermore, both CD44 and CD133 were highly expressed in sphere-forming cells vs. differentiated cells. sphere-forming cells displayed higher chemoresistance to carboplatin as opposed to differentiated cells. Moreover, intraocular injection of as few as 2x10 3 sphere-forming cells into NOD/sCID mice gave rise to new tumors similar to the original patient tumors. these results revealed that the sphere-forming cells preserved their stem cell properties and tumorigenicity, even after longterm culture. this would be a suitable in vitro model to study cancer stem-like cells in retinoblastoma and to develop chemotherapeutic drugs and strategies.

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Section: Discussionmentioning

confidence: 99%

Maintenance of retinal cancer stem cell-like properties through long-term serum-free culture from human retinoblastoma

Jiao

2011

Oncol Rep

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“…Cells passed through a 100-mm strainer (Falcon, Oxnard, Calif., U.S.A.) were then seeded (1 × 10 5 /ml) into Falcon culture flasks and grown in medium containing Dulbecco's modified Eagle's medium (DMEM)-F12 (GIBCO-Invitrogen), penicillin G, streptomycin sulfate, B-27 (GIBCO-Invitrogen), recombinant human fibroblast growth factor-2 (20 ng/ml; R&D Systems, Minneapolis, Minn., U.S.A.), and recombinant human epidermal growth factor (20 ng/ml; R&D Systems). 4,6,8,15) The medium was replaced every 3 days with identical fresh medium. Spheres were counted under a phase-contrast microscope (Olympus IMT-2, Tokyo).…”

Section: Methodsmentioning

confidence: 99%

“…To investigate tumorsphere self-renewal capacity, modified limiting dilution assays were performed. 4,8,15) Briefly, dissociated cells ware plated in 96-well plates (Falcon) in 200 ml volumes of DMEM-F12 medium with growth factors. Cultures were fed 20 ml of medium with growth factors every 2 days and the percentage of wells without tumorspheres was calculated and plotted against the number of cells per well.…”

Section: Methodsmentioning

confidence: 99%

“…Tumor samples were fixed in 4% paraformalde- Immunocytochemistry of tumorspheres was carried out as described previously 4,6,8,15) Our experimental procedures involving animals followed the guidelines of the Animal Experimental Committee of Gifu University. Tumorigenicity was determined by orthotopically injecting CSCs derived from the brain tumor as described previously.…”

Section: Methodsmentioning

confidence: 99%

“…Tumorigenicity was determined by orthotopically injecting CSCs derived from the brain tumor as described previously. 4,8,15) After 20 weeks of primary culture, cells were injected into the right flank of non-obese, diabetic, severe combined immunodeficiency mice and 2 ml of 1 × 10 8 cells/ml cell suspension in proliferation medium (total 10 4 cells) was delivered into the right striatum (0.2 ml/min) by stereotactic injection through a Hamilton syringe, all under general anesthesia. The mice were sacrificed at 12-14 weeks post-injection depending on the injected cell line.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Brain Tumor Stem Cells From an Adenoid Glioblastoma Multiforme

Oka

Soeda

Noda

et al. 2009

Neurol. Med. Chir.(Tokyo)

Self Cite

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Brain tumors may arise from and contain cancer stem cells (CSCs) capable of self-renewal, proliferation, and differentiation that recapitulate the parent tumor. These CSCs are thought to be important in gliomagenesis. Detection of CSCs invading the adjacent brain regions is important for the diagnosis and effective treatment of glioblastoma multiforme (GBM). A 57-year-old man presented with an adenoid GBM, and underwent resection of the tumor. Multipotent, self-renewing cells derived from the human adenoid GBM were isolated and identified with the tumor-derived stem cell surface antigen CD133 from whole autopsied patient's brain. Tumorsphere culture and flow cytometric analysis revealed that 1.02-2.32% of the cells were positive for CD133. Transplantation of cultured tumorspheres into the mice brain resulted in the formation of well-defined tumor masses after 12 weeks. The histological and immunohistochemical characteristics of the xenograft were identical to those of the parent tumor. Examination of the patient's brain at autopsy showed CD133-positive cells were identified in the brain regions adjacent to the tumor, suggesting that CD133-positive CSCs might be localized to the vascular niche. Methods to localize CSCs may open new approaches for the treatment of brain tumors.

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Hypoxia‐inducible factor 1α inhibits the fibroblast‐like markers type I and type III collagen during hypoxia‐induced chondrocyte redifferentiation: Hypoxia not only induces type II collagen and aggrecan, but it also inhibits type I and type III collagen in the hypoxia‐inducible factor 1α–dependent redifferentiation of chondrocytes

Duval¹,

Leclercq²,

Elissalde³

et al. 2009

Arthritis & Rheumatism

137

127

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Objective. Autologous chondrocyte implantation requires expansion of cells ex vivo, leading to dedifferentiation of chondrocytes (loss of aggrecan and type II collagen to the profit of type I and type III collagens). Several approaches have been described for redifferentiation of these cells. Among them, low oxygen tension has been exploited to restore the differentiated chondrocyte phenotype, but molecular mechanisms of this process remain unclear. However, under conditions of hypoxia, one of the major factors involved is hypoxiainducible factor 1␣ (HIF-1␣). The purpose of this study was to investigate the role of HIF-1␣ during human chondrocyte redifferentiation.Methods. We used complementary approaches to achieving HIF-1␣ loss (inhibition by cadmium ions and dominant-negative expression) or gain (ectopic expression and cobalt ion treatment) of function. Expression of chondrocyte, as well as fibroblast-like, phenotype markers was determined using real-time reverse transcription-polymerase chain reaction and Western blot analyses. Binding activities of HIF-1␣ and SOX9, a pivotal transcription factor of chondrogenesis, were evaluated by electrophoretic mobility shift assays and by chromatin immunoprecipitation assay.Results. We found that hypoxia and HIF-1␣ not only induced the expression of SOX9, COL2A1, and aggrecan, but they simultaneously inhibited the expression of COL1A1, COL1A2, and COL3A1. In addition, we identified the binding of HIF-1␣ to the aggrecan promoter, the first such reported demonstration of this binding.Conclusion. This study is the first to show a bimodal role of HIF-1␣ in cartilage homeostasis, since HIF-1␣ was shown to favor specific markers and to impair dedifferentiation. This suggests that manipulation of HIF-1␣ could represent a promising approach to the treatment of osteoarthritis.

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Long-term maintenance of brain tumor stem cell properties under at non-adherent and adherent culture conditions

Cited by 45 publications

References 18 publications

Maintenance of retinal cancer stem cell-like properties through long-term serum-free culture from human retinoblastoma

Maintenance of retinal cancer stem cell-like properties through long-term serum-free culture from human retinoblastoma

Brain Tumor Stem Cells From an Adenoid Glioblastoma Multiforme

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