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.

scholarly journals Expansive growth of two glioblastoma stem-like cell lines is mediated by bFGF and not by EGF

2013 ◽  
Vol 47 (4) ◽  
pp. 330-337 ◽  
Author(s):  
Neza Podergajs ◽  
Narve Brekka ◽  
Bernhard Radlwimmer ◽  
Christel Herold-Mende ◽  
Krishna M. Talasila ◽  
...  
Keyword(s):  
Stem Cell ◽  
Cell Lines ◽  
Basal Medium ◽  
Growth Stimulation ◽  
Growth Conditions ◽  
Stem Cell Markers ◽  
Egfr Amplification ◽  
Egfr Expression ◽  
Stem Cell Medium ◽  
Cell Medium

Abstract Background. Patient-derived glioblastoma (GBM) stem-like cells (GSCs) represent a valuable model for basic and therapeutic research. GSCs are usually propagated in serum-free Neural Basal medium supplemented with bFGF and EGF. Yet, the exact influence of these growth factors on GSCs is still unclear. Recently it was suggested that GBM stemlike cells with amplified EGFR should be cultured in stem cell medium without EGF, as the presence of EGF induced rapid loss of EGFR amplification. However, patient biopsies are usually taken into culture before their genomic profiles are defined. Thus, an important question remains whether GBM cells without EGFR amplification also can be cultured in stem cell medium without EGF. Meterials and methods. To address this question, we used two heterogeneous glioblastoma GSC lines (NCH421k and NCH644) that lack EGFR amplification. Results. Although both cell lines showed very low EGFR expression under standard growth conditions, bFGF stimulation induced higher expression of EGFR in NCH644. In both cell lines, expression of the stem cell markers nestin and CD133 was higher upon stimulation with bFGF compared to EGF. Importantly, bFGF stimulated the growth of both cell lines, whereas EGF had no effect. We verified that the growth stimulation by bFGF was either mediated by proliferation (NCH421k) or resistance to apoptosis (NCH644). Conclusions. We demonstrate that GSC cultures without EGFR amplification can be maintained and expanded with bFGF, while the addition of EGF has no significant effect and therefore can be omitted.

397 DERIVATION OF NEURAL STEM CELLS FROM PORCINE EPIBLAST CELLS

2010 ◽  
Vol 22 (1) ◽  
pp. 355
Author(s):  
M. A. Rasmussen ◽  
V. J. Hall ◽  
P. Hyttel
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Neural Stem Cells ◽  
Neural Stem Cell ◽  
Bipolar Cells ◽  
Stem Cell Markers ◽  
High Density Culture ◽  
Cell Markers ◽  
Stem Cell Medium ◽  
Cell Medium

The use of neural stem cells (NSC) has gained increased attention as a means of treating neurodegenerative diseases such as Alzheimer’s and Parkinson’s disease. However, before regenerative treatment of humans can be undertaken, safety studies of NSC using animal models are required. The mouse has been the model of choice so far; however, testing in larger mammals such as the pig is essential. The aim of this study was to derive NSC from porcine epiblast cells and to analyze these cells using neural stem cell markers. A total of 47 epiblasts were isolated from E9 porcine embryos and grown on mouse embryonic fibroblast cells in a porcine embryonic stem cell medium. After 5 days, 23 outgrowth colonies had formed (49%). Based on morphology, 8 outgrowth colonies were selected and cut into 63 smaller pieces, which were transferred to MS5 stromal cells in a serum replacement medium, and after an additional 12 days, rosette structures had formed. These structures were transferred to Matrigel-coated dishes in a neural stem cell medium containing EGF and FGF. Under such conditions, bipolar cells containing large nuclei and several nucleoli grew out from the rosettes. The bipolar cells have been expanded for more than 8 passages without any change in morphology or growth rate, and upon high-density culture, the cells spontaneously form floating neurospheres. Stainings revealed that the cells expressed the neural stem cell markers Nestin (100%), Sox2 (100%), Pax6 (100%), and Vimentin (100%), as well as the proliferation marker Ki67 (54%). The same markers were found to be expressed in the lateral ventricles of the developing porcine brain, a location known to have high neurogenic activity. When growth factors were withdrawn from the culture medium, a higher proportion of TujI expressing cells were observed, especially when cells were cultured as neurospheres. We conclude that it is possible to derive presumptive NSC from porcine epiblast cells and that these express the same markers as reported for human NSC. Further studies are required to determine if the cells can be cultured long term and differentiate into various neuronal and glial cell types.

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

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 Hypoxic-Preconditioned Bone Marrow Stem Cell Medium Significantly Improves Outcome After Retinal Ischemia in Rats

2016 ◽  
Vol 57 (7) ◽  
pp. 3522 ◽  
Author(s):  
Steven Roth ◽  
John C. Dreixler ◽  
Biji Mathew ◽  
Irina Balyasnikova ◽  
Jacob R. Mann ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stem Cell ◽  
Retinal Ischemia ◽  
Bone Marrow Stem Cell ◽  
Stem Cell Medium ◽  
Cell Medium ◽  
Marrow Stem Cell

scholarly journals Direct reprogramming of epithelial cell rests of malassez into mesenchymal-like cells by epigenetic agents

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Koki Yoshida ◽  
Osamu Uehara ◽  
Yoshihito Kurashige ◽  
Durga Paudel ◽  
Aya Onishi ◽  
...  
Keyword(s):  
Stem Cell ◽  
Epithelial Cell ◽  
Periodontal Regeneration ◽  
Embryonic Stem ◽  
Stem Cell Markers ◽  
Cell Markers ◽  
Demethylating Agent ◽  
Protein Levels ◽  
Deacetylase Inhibitor ◽  
Stem Cell Medium

AbstractThe DNA demethylating agent, 5-Azacytidine (5Aza), and histone deacetylase inhibitor, valproic acid (Vpa), can improve the reprogramming efficiencies of pluripotent cells. This study aimed to examine the roles of 5Aza and Vpa in the dedifferentiation of epithelial cell rests of Malassez (ERM) into stem-like cells. Additionally, the ability of stem-like cells to differentiate into mesenchymal cells was evaluated. ERM was cultured in embryonic stem cell medium (ESCM) with 1 µM of 5Aza, or 2 mM of Vpa, or a combination of 5Aza and Vpa. The cells stimulated with both 5Aza and Vpa were named as progenitor-dedifferentiated into stem-like cells (Pro-DSLCs). The Pro-DSLCs cultured in ESCM alone for another week were named as DSLCs. The stem cell markers were significantly higher in the DSLCs than the controls (no additions). The mRNA and protein levels of the endothelial, mesenchymal stem, and osteogenic cell markers were significantly higher in the Pro-DSLCs and DSLCs than the controls. The combination of a demethylating agent and a deacetylated inhibitor induced the dedifferentiation of ERM into DSLCs. The Pro-DSLCs derived from ERM can be directly reprogrammed into mesenchymal-like cells without dedifferentiation into stem-like cells. Isolated ERM treated with epigenetic agents may be used for periodontal regeneration.

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.

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