New strategy for the analysis of phenotypic marker antigens in brain tumor–derived neurospheres in mice and humans

2008 ◽  
Vol 24 (3-4) ◽  
pp. E28 ◽  
Author(s):  
Anne-Marie Bleau ◽  
Brian M. Howard ◽  
Lauren A. Taylor ◽  
Demirkan Gursel ◽  
Jeffrey P. Greenfield ◽  
...  
Keyword(s):  
Stem Cell ◽  
Brain Tumor ◽  
Rapid Assessment ◽  
Solid Sphere ◽  
Phenotypic Marker ◽  
New Strategy ◽  
Stem Cell Medium ◽  
Brain Tumor Stem Cells ◽  
Cell Medium

Object Brain tumor stem cells (TSCs) hypothetically drive the malignant phenotype of glioblastoma multiforme (GBM), and evidence suggests that a better understanding of these TSCs will have profound implications for treating gliomas. When grown in vitro, putative TSCs grow as a solid sphere, making their subsequent characterization, particularly the cells within the center of the sphere, difficult. Therefore, the purpose of this study was to develop a new method to better understand the proteomic profile of the entire population of cells within a sphere. Methods Tumor specimens from patients with confirmed GBM and glioma models in mice were mechanically and enzymatically dissociated and grown in traditional stem cell medium to generate neurospheres. The neurospheres were then embedded in freezing medium, cryosectioned, and analyzed with immunofluorescence. Results By sectioning neurospheres as thinly as 5 μm, the authors overcame many of the problems associated with immunolabeling whole neurospheres, such as antibody penetration into the core of the sphere and intense background fluorescence that obscures the specificity of immunoreactivity. Moreover, the small quantity of material required and the speed with which this cryosectioning and immunolabeling technique can be performed make it an attractive tool for the rapid assessment of TSC character. Conclusions This study is the first to show that cryosectioning of neurospheres derived from glioma models in mice and GBM in humans is a feasible method of better defining the stem cell profile of a glioma.

Generation of large pig and bovine blastocysts by culturing in human induced pluripotent stem cell medium

Zygote ◽  
2015 ◽  
Vol 24 (2) ◽  
pp. 236-244 ◽  
Author(s):  
Qing-Shan Gao ◽  
Long Jin ◽  
Suo Li ◽  
Hai-Ying Zhu ◽  
Qing Guo ◽  
...  
Keyword(s):  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
State University ◽  
North Carolina State University ◽  
Stem Cell Medium ◽  
Induced Pluripotent ◽  
Cell Medium

SummaryWe investigated the effect of human induced pluripotent stem cell (hiPS) medium on porcine somatic cell nuclear transfer and bovine in vitro fertilized early blastocysts, in comparison with North Carolina State University (NCSU)-37 medium and in vitro culture (IVC)-II medium. After 2 days of culture, the diameter of the portion of the blastocyst that was extruded from the zona pellucid dramatically differed between porcine blastocysts cultured in hiPS medium and those cultured in NCSU-37 medium (221.47 ± 38.94 μm versus 481.87 ± 40.61 μm, P < 0.01). Moreover, the diameter of the portion of the blastocyst significantly differed between bovine blastocysts cultured in hiPS medium and those cultured in IVC-II medium (150.30 ± 29.49 μm versus 195.58 ± 41.59 μm, P < 0.01). Furthermore, the total number of cells per porcine and bovine blastocyst was more than two-fold higher in blastocysts cultured in hiPS medium than in those cultured in NCSU-37 medium (44.33 ± 5.28 and 143.33 ± 16.05, P < 0.01) or IVC-II medium (172.12 ± 45.08 and 604.83 ± 242.64, P < 0.01), respectively. These results indicate that hiPS medium markedly improves the quality of porcine and bovine blastocysts.

scholarly journals Osteoclastogenic Potential of Tissue-Engineered Periosteal Sheet: Effects of Culture Media on the Ability to Recruit Osteoclast Precursors

2021 ◽  
Vol 22 (4) ◽  
pp. 2169
Author(s):  
Kohya Uematsu ◽  
Takashi Ushiki ◽  
Hajime Ishiguro ◽  
Riuko Ohashi ◽  
Suguru Tamura ◽  
...  
Keyword(s):  
Stem Cell ◽  
Fluorescent Protein ◽  
Bone Marrow Cells ◽  
Culture Media ◽  
Γ Irradiation ◽  
Stem Cell Medium ◽  
Cell Medium ◽  
Conventional Medium

Cell culture media influence the characteristics of human osteogenic periosteal sheets. We have previously found that a stem cell medium facilitates growth and collagen matrix formation in vitro and osteogenesis in vivo. However, it has not yet been demonstrated which culture medium is superior for osteoclastogenesis, a prerequisite for reconstruction of normal bone metabolic basis. To address this question, we compared chemotaxis and osteoclastogenesis in tissue-engineered periosteal sheets (TPSs) prepared with two types of culture media. Periosteal tissues obtained from adult volunteers were expanded with the conventional Medium 199 or with the stem cell medium, MesenPRO. Hematopoietic enhanced-green-fluorescent-protein (EGFP)-nude mice were prepared by γ-irradiation of Balb/c nu/nu mice and subsequent transplantation of bone marrow cells from CAG-EGFP C57BL/6 mice. TPSs were implanted subcutaneously into the chimeric mice and retrieved after intervals for immunohistopathological examination. EGFP+ cells were similarly recruited to the implantation site in both the TPSs prepared, whereas the distribution of CD11b+ cells was significantly lower in the TPS prepared with the stem cell medium. Instead, osteoclastogenesis was higher in the TPS prepared with the stem cell medium than in the one prepared with the conventional medium. These findings suggest that the stem cell medium is preferable for the preparation of more functional TPSs.

Brain Tumor Stem Cells: Bringing Order to the Chaos of Brain Cancer

2008 ◽  
Vol 26 (17) ◽  
pp. 2916-2924 ◽  
Author(s):  
Peter B. Dirks
Keyword(s):  
Stem Cell ◽  
Brain Tumor ◽  
Cell Biology ◽  
In Vivo Models ◽  
Brain Tumor Cells ◽  
Brain Tumor Stem Cells ◽  
New Treatments ◽  
Key Pathways

Brain tumors are generally incurable cancers. Work from a number of laboratories strongly suggests that they are organized as a hierarchy based on a subset of cancer cells that have stem-cell properties. These cells have now been shown to be resistant to conventional therapy and responsive to differentiation therapy. New in vitro and in vivo models for interrogating brain tumor cells in stem-cell conditions have been developed that provide important new opportunities for elucidating the key pathways responsible for driving the proliferation of these cells. Continued application of the principles of stem-cell biology to the study of brain cancers is likely to continue to bring further important insight into these aggressive cancers, bringing new treatments and understanding of the origins.

341 FELINE EMBRYONIC STEM-LIKE CELLS DERIVED FROM IN VITRO-PRODUCED BLASTOCYSTS RETAIN IN VITRO DIFFERENTIATION POTENTIAL

2015 ◽  
Vol 27 (1) ◽  
pp. 259
Author(s):  
T. Tharasanit ◽  
N. Tiptanavattana ◽  
P. Phakdeedindan ◽  
M. Techakumphu
Keyword(s):  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Cell Lines ◽  
Es Cells ◽  
Embryonic Stem ◽  
In Vitro Differentiation ◽  
Stem Cell Medium ◽  
Cell Medium ◽  
Embryonic Stem Cell Medium

Embryonic stem (ES) cells are pluripotent cells that can differentiate into all 3 germ layers, including endoderm, mesoderm, and ectoderm. Embryonic stem cells are generally divided into 2 types, naïve and primed-state, depending on their signaling pathways. Domestic cat is a useful animal model for the study of human diseases because many genetic and infectious diseases in the cat are analogous with similar aetiology to human diseases. The cat can also be used as a research model for reproductive physiology and conservation of wild felids. Until recently, information on establishment of feline ES cells is limited. The objectives of this study were to isolate cat ES cells from in vitro-produced blastocysts and to examine the effect of different concentrations of basic fibroblast growth factor (bFGF) on the expression of pluripotent genes. Inner cell masses (ICM) from cat blastocysts (n = 40, Day 7 after in vitro fertilization) that were matured, fertilized, and cultured entirely in vitro, were isolated by immunosurgery and plated on mitmycin-treated mouse embryonic fibroblasts. The ICM (n = 20) were then cultured in embryonic stem cell medium containing 1000 IU mL–1 of leukemia inhibitory factor (LIF) and different bFGF concentrations (0, 4, 10, and 20 ng mL–1). The ICM outgrowths at 7 days postplating were collected and analysed for expression of pluripotent genes (SOX-2, OCT-4, and NANOG). Results showed that transcription levels of all 3 pluripotent genes were higher in ICM outgrowths cultured in 20 ng mL–1 of bFGF compared with the lower concentrations. For isolation of ES cells, ICM (n = 20) were cultured in embryonic stem cell medium supplemented with 1000 IU mL–1 of LIF and 20 ng mL–1 of bFGF due to the results obtained from the above experiment. Established ES cells were characterised by detecting alkaline phosphatase (AP) activity and expression of ES markers (SOX-2, OCT-4, SSEA-4) at protein level, and karyotyped at passage 20 and 40. In vitro differentiation into embryoid bodies (EB) was induced by the hanging drop technique, and EB samples (n = 5 for each time point) were tested for the expression of TTR, AFP, T (Bracyury), NKX2.5, MAP-2, and NESTIN genes at 0, 7, and 14 days of culture. A total of 3 ES-like cell lines were established with a typical ES morphology, such as a well-defined colony, a large nucleus to cytoplasm ratio with 1 to 2 prominent nucleoli. The 3 ES-like cell lines were passaged up to 40 times with a normal diploid karyotype (n = 38). They were strongly positive for AP, SOX-2, OCT-4, and SSEA-4. Following EB culture, cell aggregation and cystic-like structure were observed. The EB samples also expressed all differentiation markers. This study reports that feline ES-like cell lines can be generated from in vitro-produced feline blastocysts. The ES cell lines can be repeatedly passaged indicating self-renewal ability, and gene expression of the EB demonstrates cellular differentiation into all 3 germ layers.

scholarly journals A stem cell medium containing neural stimulating factor induces a pancreatic cancer stem-like cell-enriched population

2014 ◽  
Vol 45 (5) ◽  
pp. 1857-1866 ◽  
Author(s):  
YUSAKU WATANABE ◽  
KIYOSHI YOSHIMURA ◽  
KOICHI YOSHIKAWA ◽  
RYOICHI TSUNEDOMI ◽  
YOSHITARO SHINDO ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
Stem Cell ◽  
Stem Cell Medium ◽  
Cell Medium ◽  
Stimulating Factor

scholarly journals Alpha 1-antichymotrypsin contributes to stem cell characteristics and enhances tumorigenicity of Glioblastoma

2020 ◽  
Author(s):  
Montserrat Lara-Velazquez ◽  
Natanael Zarco ◽  
Anna Carrano ◽  
Jordan Phillipps ◽  
Emily S Norton ◽  
...  
Keyword(s):  
Stem Cell ◽  
Cell Migration ◽  
Brain Tumor ◽  
Primary Cultures ◽  
Cellular Migration ◽  
Tumor Initiating Cells ◽  
The Impact

Abstract Background Glioblastomas (GBMs) are the most common primary brains tumors in adults with almost 100% recurrence rate. Patients with lateral ventricle proximal GBMs (LV-GBMs) exhibit worse survival compared to distal locations for reasons that remain unknown. One potential explanation is the proximity of these tumors to the cerebrospinal fluid (CSF) and its contained chemical cues that can regulate cellular migration and differentiation. We therefore investigated the role of CSF on GBM gene expression and the role of a CSF-induced gene, SERPINA3, in GBM malignancy in vitro and in vivo. Methods We utilized patient-derived CSF and primary cultures of GBM brain tumor initiating cells (BTICs). We determined the impact of SERPINA3 expression in glioma patients using TCGA database. SERPINA3 expression changes were evaluated at both the mRNA and protein levels. The effects of knockdown (KD) and overexpression (OE) of SERPINA3 on cell behavior were evaluated by transwell assay (for cell migration), and alamar blue and Ki67 (for viability and proliferation respectively). Stem cell characteristics on KD cells were evaluated by differentiation and colony formation experiments. Tumor growth was studied by intracranial and flank injections. Results GBM CSF induced a significant increase in BTIC migration accompanied by upregulation of the SERPINA3 gene. In patient samples and TCGA data we observed SERPINA3 to correlate directly with brain tumor grade and indirectly with GBM patient survival. Silencing of SERPINA3 induced a decrease in cell proliferation, migration, invasion, and stem cell characteristics, while SERPINA3 overexpression increased cell migration. In vivo, mice orthotopically-injected with SERPINA3 KD BTICs showed increased survival. Conclusions SERPINA3 plays a key role in GBM malignancy and its inhibition results in a better outcome using GBM preclinical models.

scholarly journals Identification of a native novel oncolytic immunoglobulin on exfoliated colon epithelial cells: A bispecific heterodimeric chimera of IgA/IgG

2017 ◽  
Author(s):  
George P. Albaugh ◽  
Sudhir K. Dutta ◽  
Vasantha Iyengar ◽  
Samina Shami ◽  
Althaf Lohani ◽  
...  
Keyword(s):  
Stem Cell ◽  
Direct Method ◽  
Stem Cell Markers ◽  
Distinct Cell ◽  
Stem Cell Medium ◽  
Stool Samples ◽  
Low Levels ◽  
Colonic Cells ◽  
Cell Medium ◽  
A Direct Method

ABSTRACTUnderstanding the nature of cell surface markers on exfoliated colonic cells is a crucial step in establishing criteria for a normally functioning mucosa. We have found that colonic cells isolated from stool samples (SCSR-010 Fecal Cell Isolation Kit, NonInvasive Technologies, Elkridge, MD), preserved at room temperature for up to one week, with viability of >85% and low levels of apoptosis (8% - 10%) exhibit two distinct cell size subpopulations, in the 2.5μM– 5.0 μM and 5.0μM-8.0μM range. In addition to IgA, about 60% of the cells expressed a novel heterodimeric IgA/IgG immunoglobulin that conferred a broad-spectrum cell mediated cytotoxicity against tumor cells. In a cohort of 58 subjects the exclusive absence of this immunoglobulin in two African-Americans was suggestive of a germline deletion. Serial cultures in stem cell medium retained the expression of this heterodimer. Since a majority of the cystic cells expressed the stem cell markers Lgr5 and Musashi-1 we termed these cells as gastrointestinal progenitor stem cells (GIP-C**). CXCR-4, the cytokine co-receptor for HIV was markedly expressed. These cells also expressed CD20, IgA, IgG, CD45, and COX-2. We assume that they originated from mature columnar epithelium by dedifferentiation. Our observations indicate that we have a robust noninvasive method to study mucosal pathophysiology and a direct method to create a database for applications in regenerative medicine.

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