7 results on '"von Furstenberg, Richard J."'
Search Results
2. CD24 can be used to isolate Lgr5+ putative colonic epithelial stem cells in mice
- Author
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King, Jeffrey B., primary, von Furstenberg, Richard J., additional, Smith, Brian J., additional, McNaughton, Kirk K., additional, Galanko, Joseph A., additional, and Henning, Susan J., additional
- Published
- 2012
- Full Text
- View/download PDF
3. Sorting mouse jejunal epithelial cells with CD24 yields a population with characteristics of intestinal stem cells
- Author
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von Furstenberg, Richard J., primary, Gulati, Ajay S., additional, Baxi, Anand, additional, Doherty, Jason M., additional, Stappenbeck, Thaddeus S., additional, Gracz, Adam D., additional, Magness, Scott T., additional, and Henning, Susan J., additional
- Published
- 2011
- Full Text
- View/download PDF
4. CD24 can be used to isolate Lgr5+ putative colonic epithelial stem cells in mice.
- Author
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King, Jeffrey B., von Furstenberg, Richard J., Smith, Brian J., McNaughton, Kirk K., Galanko, Joseph A., and Henning, Susan J.
- Abstract
A growing body of evidence has implicated CD24, a cell-surface protein, as a marker of colorectal cancer stem cells and target for antitumor therapy, although its presence in normal colonic epithelium has not been fully characterized. Previously, our group showed that CD24-based cell sorting can be used to isolate a fraction of murine small intestinal epithelial cells enriched in actively cycling stem cells. Similarly, we hypothesized that CD24-based isolation of colonic epithelial cells would generate a fraction enriched in actively cycling colonic epithelial stem cells (CESCs). Immunohistochemistry performed on mouse colonic tissue showed CD24 expression in the bottom half of proximal colon crypts and the crypt base in the distal colon. This pattern of distribution was similar to enhanced green fluorescent protein (EGFP) expression in Lgr5-EGFP mice. Areas expressing CD24 contained actively proliferating cells as determined by ethynyl deoxyuridine (EdU) incorporation, with a distinct difference between the proximal colon, where EdU-labeled cells were most frequent in the midcrypt, and the distal colon, where they were primarily at the crypt base. Flow cytometric analyses of single epithelial cells, identified by epithelial cell adhesion molecule (EpCAM) positivity, from mouse colon revealed an actively cycling CD24+ fraction that contained the majority of Lgr5-EGFP+putative CESCs. Transcript analysis by quantitative RT-PCR confirmed enrichment of active CESC markers [leucine-rich-repeat-containing G protein-coupled receptor 5 (Lgr5), ephrin type B receptor 2 (EphB2), and CD166] in the CD24+EpCAM+ fraction but also showed enrichment of quiescent CESC markers [leucine-rich repeats and immunoglobin domains (Lrig), doublecortin and calmodulin kinase- like 1 (DCAMKL-1), and murine telomerase reverse transcriptase (mTert)]. We conclude that CD24-based sorting in wild-type mice isolates a colonic epithelial fraction highly enriched in actively cycling and quiescent putative CESCs. Furthermore, the presence of CD24 expression in normal colonic epithelium may have important implications for the use of anti-CD24-based colorectal cancer therapies. [ABSTRACT FROM AUTHOR]
- Published
- 2012
- Full Text
- View/download PDF
5. Ductular and proliferative response of esophageal submucosal glands in a porcine model of esophageal injury and repair.
- Author
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Krüger L, Gonzalez LM, Pridgen TA, McCall SJ, von Furstenberg RJ, Harnden I, Carnighan GE, Cox AM, Blikslager AT, and Garman KS
- Subjects
- Active Transport, Cell Nucleus, Animals, Esophageal Diseases pathology, Female, Gene Expression Regulation physiology, Humans, Male, SOX9 Transcription Factor genetics, SOX9 Transcription Factor metabolism, Staining and Labeling, Swine, Cell Proliferation physiology, Esophagus cytology, Esophagus injuries
- Abstract
Esophageal injury is a risk factor for diseases such as Barrett's esophagus (BE) and esophageal adenocarcinoma. To improve understanding of signaling pathways associated with both normal and abnormal repair, animal models are needed. Traditional rodent models of esophageal repair are limited by the absence of esophageal submucosal glands (ESMGs), which are present in the human esophagus. Previously, we identified acinar ductal metaplasia in human ESMGs in association with both esophageal injury and cancer. In addition, the SOX9 transcription factor has been associated with generation of columnar epithelium and the pathogenesis of BE and is present in ESMGs. To test our hypothesis that ESMGs activate after esophageal injury with an increase in proliferation, generation of a ductal phenotype, and expression of SOX9, we developed a porcine model of esophageal injury and repair using radiofrequency ablation (RFA). The porcine esophagus contains ESMGs, and RFA produces a consistent and reproducible mucosal injury in the esophagus. Here we present a temporal assessment of this model of esophageal repair. Porcine esophagus was evaluated at 0, 6, 18, 24, 48, and 72 h and 5 and 7 days following RFA and compared with control uninjured esophagus. Following RFA, ESMGs demonstrated an increase in ductal phenotype, echoing our prior studies in humans. Proliferation increased in both squamous epithelium and ESMGs postinjury with a prominent population of SOX9-positive cells in ESMGs postinjury. This model promises to be useful in future experiments evaluating mechanisms of esophageal repair. NEW & NOTEWORTHY A novel porcine model of injury and repair using radiofrequency ablation has been developed, allowing for reproducible injury to the esophagus to study repair in an animal model with esophageal submucosal glands, a key anatomical feature and missing in rodent models but possibly harboring progenitor cells. There is a strong translational component to this porcine model given the anatomical and physiological similarities between pigs and humans., (Copyright © 2017 the American Physiological Society.)
- Published
- 2017
- Full Text
- View/download PDF
6. CD24 can be used to isolate Lgr5+ putative colonic epithelial stem cells in mice.
- Author
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King JB, von Furstenberg RJ, Smith BJ, McNaughton KK, Galanko JA, and Henning SJ
- Subjects
- Animals, Antigens, Neoplasm metabolism, Biomarkers metabolism, Cell Adhesion Molecules metabolism, Cell Proliferation, Colon cytology, Epithelial Cell Adhesion Molecule, Gene Expression Regulation, Green Fluorescent Proteins biosynthesis, Green Fluorescent Proteins genetics, Immunohistochemistry, Intestinal Mucosa cytology, Mice, Mice, Inbred C57BL, Mice, Transgenic, Microscopy, Confocal, RNA, Messenger metabolism, Receptors, G-Protein-Coupled genetics, Reverse Transcriptase Polymerase Chain Reaction, Transcription, Genetic, CD24 Antigen metabolism, Cell Separation methods, Colon immunology, Epithelial Cells immunology, Flow Cytometry, Intestinal Mucosa immunology, Receptors, G-Protein-Coupled metabolism, Stem Cells immunology
- Abstract
A growing body of evidence has implicated CD24, a cell-surface protein, as a marker of colorectal cancer stem cells and target for antitumor therapy, although its presence in normal colonic epithelium has not been fully characterized. Previously, our group showed that CD24-based cell sorting can be used to isolate a fraction of murine small intestinal epithelial cells enriched in actively cycling stem cells. Similarly, we hypothesized that CD24-based isolation of colonic epithelial cells would generate a fraction enriched in actively cycling colonic epithelial stem cells (CESCs). Immunohistochemistry performed on mouse colonic tissue showed CD24 expression in the bottom half of proximal colon crypts and the crypt base in the distal colon. This pattern of distribution was similar to enhanced green fluorescent protein (EGFP) expression in Lgr5-EGFP mice. Areas expressing CD24 contained actively proliferating cells as determined by ethynyl deoxyuridine (EdU) incorporation, with a distinct difference between the proximal colon, where EdU-labeled cells were most frequent in the midcrypt, and the distal colon, where they were primarily at the crypt base. Flow cytometric analyses of single epithelial cells, identified by epithelial cell adhesion molecule (EpCAM) positivity, from mouse colon revealed an actively cycling CD24(+) fraction that contained the majority of Lgr5-EGFP(+) putative CESCs. Transcript analysis by quantitative RT-PCR confirmed enrichment of active CESC markers [leucine-rich-repeat-containing G protein-coupled receptor 5 (Lgr5), ephrin type B receptor 2 (EphB2), and CD166] in the CD24(+)EpCAM(+) fraction but also showed enrichment of quiescent CESC markers [leucine-rich repeats and immunoglobin domains (Lrig), doublecortin and calmodulin kinase-like 1 (DCAMKL-1), and murine telomerase reverse transcriptase (mTert)]. We conclude that CD24-based sorting in wild-type mice isolates a colonic epithelial fraction highly enriched in actively cycling and quiescent putative CESCs. Furthermore, the presence of CD24 expression in normal colonic epithelium may have important implications for the use of anti-CD24-based colorectal cancer therapies.
- Published
- 2012
- Full Text
- View/download PDF
7. Sorting mouse jejunal epithelial cells with CD24 yields a population with characteristics of intestinal stem cells.
- Author
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von Furstenberg RJ, Gulati AS, Baxi A, Doherty JM, Stappenbeck TS, Gracz AD, Magness ST, and Henning SJ
- Subjects
- Animals, Biomarkers metabolism, CD24 Antigen genetics, Cell Adhesion Molecules metabolism, Cell Proliferation, Cells, Cultured, Doublecortin-Like Kinases, Epithelial Cells metabolism, Gene Expression Profiling methods, Gene Expression Regulation, Immunohistochemistry, Jejunum cytology, Jejunum metabolism, Leukocyte Common Antigens deficiency, Male, Mice, Mice, Inbred C57BL, Mice, Transgenic, Nuclear Proteins metabolism, Oligonucleotide Array Sequence Analysis, Paneth Cells metabolism, Platelet Endothelial Cell Adhesion Molecule-1 metabolism, Polycomb Repressive Complex 1, Protein Serine-Threonine Kinases metabolism, Proto-Oncogene Proteins metabolism, RNA, Messenger metabolism, Receptors, G-Protein-Coupled genetics, Receptors, G-Protein-Coupled metabolism, Repressor Proteins metabolism, Reverse Transcriptase Polymerase Chain Reaction, Stem Cells metabolism, Time Factors, CD24 Antigen metabolism, Cell Separation methods, Epithelial Cells immunology, Flow Cytometry, Jejunum immunology, Paneth Cells immunology, Stem Cells immunology
- Abstract
Intestinal stem cells (ISCs) have been studied for more than three decades; however, their isolation has remained a challenge. We hypothesized that, just as for stem cells of other tissues, one or more membrane markers would allow positive selection of ISCs by antibody-based sorting. To explore this hypothesis, microarray data of putative ISC fractions generated by side population sorting and laser capture microdissection were subjected to bioinformatic analysis to identify common membrane antigens. The microarray comparison suggested CD24 as a candidate surface marker, and immunohistochemistry showed expression of CD24 in epithelial cells of crypt bases. Flow cytometry of jejunal epithelial preparations revealed a CD24(+) CD45(-) fraction comprising ∼1% of the cells. Analysis with epithelial cell adhesion molecule and CD31 confirmed that the cell preparations were epithelial and without endothelial contamination. Cycling cells identified by prior injection with 5-ethynyl-2'-deoxyuridine were found predominantly in the CD24(lo) subfraction. Transcript analysis by real-time RT-PCR showed this subfraction to be enriched in the ISC markers leucine-rich-repeat-containing G-protein-coupled receptor 5 (40-fold) and Bmi1 (5-fold), but also enriched in lysozyme (10-fold). Flow cytometry with anti-lysozyme antibodies demonstrated that Paneth cells comprise ∼30% of the CD24(lo) subfraction. Additional flow analyses with leucine-rich-repeat-containing G-protein-coupled receptor 5-enhanced green fluorescent protein (EGFP) epithelium demonstrated colocalization of EGFP(hi) and CD24(lo). In contrast, CD24 cells were negative for the quiescent ISC marker doublecortin and CaM kinase-like-1. Culture of CD24(lo) cells in Matrigel generated organoid structures, which included all four epithelial lineages, thus giving functional evidence for the presence of ISCs. We conclude that the CD24(lo) fraction of jejunal epithelium is highly enriched with cycling ISCs. This isolation method should be useful to many investigators in the field to advance both the basic understanding of ISC biology and the therapeutic applications of ISCs.
- Published
- 2011
- Full Text
- View/download PDF
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