The Experts below are selected from a list of 7578 Experts worldwide ranked by ideXlab platform
Robert M Hoffman - One of the best experts on this subject based on the ideXlab platform.
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Hair Follicle-Associated Pluripotent (HAP) Stem Cells in Gelfoam® Histoculture for Use in Spinal Cord Repair.
3D Sponge-Matrix Histoculture, 2018Co-Authors: Fang Liu, Robert M HoffmanAbstract:The stem cell marker, nestin, is expressed in the hair follicle, both in cells in the bulge area (BA) and the dermal papilla (DP). Nestin-expressing hair follicle-associated-pluripotent (HAP) stem cells of both the BA and DP have been previously shown to be able to form neurons, heart muscle cells, and other non-follicle cell types. The ability of the nestin-expressing HAP stem cells from the BA and DP to repair spinal cord injury was compared. Nestin-expressing HAP stem cells from both the BA and DP grew very well on Gelfoam®. The HAP stem cells attached to the Gelfoam® within 1 h. They grew along the grids of the Gelfoam® during the first 2 or 3 days. Later they spread into the Gelfoam®. After transplantation of Gelfoam® cultures of nestin-expressing BA or DP HAP stem cells into the injured spinal cord (including the Gelfoam®) nestin-expressing BA and DP cells were observed to be viable over 100 days post-surgery. Hematoxylin and eosin (H&E) staining showed connections between the transplanted cells and the host spine tissue. Immunohistochemistry showed many Tuj1-, Isl 1/2, and EN1-positive cells and nerve fibers in the transplanted area of the spinal cord after BA Gelfoam® or DP Gelfoam® cultures were transplanted to the spine. The spinal cord of mice was injured to effect hind-limb paralysis. Twenty-eight days after transplantation with BA or DP HAP stem cells on Gelfoam® to the injured area of the spine, the mice recovered normal locomotion.
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Nerve Growth and Interaction in Gelfoam® Histoculture: A Nervous System Organoid.
3D Sponge-Matrix Histoculture, 2018Co-Authors: Robert M Hoffman, Sumiyuki Mii, Jennifer Duong, Yasuyuki AmohAbstract:Nestin-expressing hair follicle-associated pluripotent (HAP) stem cells reside mainly in the bulge area (BA) of the hair follicle but also in the dermal papilla (DP). The BA appears to be origin of HAP stem cells. Long-term Gelfoam® histoculture was established of whiskers isolated from transgenic mice, in which there is nestin-driven green fluorescent protein (ND-GFP). HAP stem cells trafficked from the BA toward the DP area and extensively grew out onto Gelfoam® forming nerve-like structures. These fibers express the neuron marker β-III tubulin-positive fibers and consisted of ND-GFP-expressing cells and extended up to 500 mm from the whisker nerve stump in Gelfoam® histoculture. The growing fibers had growth cones on their tips expressing F-actin indicating that the fibers were growing axons. HAP stem cell proliferation resulted in elongation of the follicle nerve and interaction with other nerves in 3D Gelfoam® histoculture, including the sciatic nerve, trigeminal nerve, and trigeminal nerve ganglion.
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Imaging the Governing Step of Metastasis in Gelfoam® Histoculture.
3D Sponge-Matrix Histoculture, 2018Co-Authors: Robert M Hoffman, Takashi ChishimaAbstract:Distant organ colonization by cancer cells is the governing step of metastasis. We review in this chapter the modeling and imaging of organ colonization by cancer cells in Gelfoam® histoculture. ANIP 973 lung cancer cells expressing green fluorescent protein (GFP) were injected intravenously into nude mice, whereby they formed brilliantly fluorescing metastatic colonies on the mouse lung. The seeded lung tissue was then excised and incubated in the three-dimensional Gelfoam® histoculture that maintained the critical features of progressive in vivo organ colonization. Tumor progression was continuously visualized by GFP fluorescence of individual cultures over a 52-day period, during which tumor colonies spread throughout the lung. Organ colonization was selective in Gelfoam® histoculture for lung cancer cells to grow on lung tissue, since no growth occurred on histocultured mouse liver tissue. The ability to support selective organ colonization in Gelfoam® histoculture and visualize tumor progression by GFP fluorescence allows the in vitro study of the governing processes of metastasis.
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Imaging DNA Repair After UV Irradiation Damage of Cancer Cells in Gelfoam® Histoculture.
3D Sponge-Matrix Histoculture, 2018Co-Authors: Shinji Miwa, Robert M HoffmanAbstract:DNA damage repair in response to UVC irradiation was imaged in cancer cells growing in Gelfoam® histoculture. UVC-induced DNA damage repair was imaged with green fluorescent protein (GFP) fused to the DNA damage response (DDR)-related binding protein 53BP1 in MiaPaCa-2 human pancreatic cancer cells. Three-dimensional Gelfoam® histocultures and confocal imaging enabled 53BP1-GFP nuclear foci to be observed within 1 h after UVC irradiation, indicating the onset of DNA damage repair response. Induction of UV-induced 53BP1-GFP focus formation was limited up to a depth of 40 μm in Gelfoam® histoculture of MiaPaCa-2 cells, indicating this was the depth limit of UVC irradiation.
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Hair-Shaft Growth in Gelfoam® Histoculture of Skin and Isolated Hair Follicles.
3D Sponge-Matrix Histoculture, 2018Co-Authors: Robert M Hoffman, Wenluo CaoAbstract:Human scalp skin with abundant hair follicles in various stages of the hair growth cycle was histocultured for up to 40 days on Gelfoam® at the air/liquid interface. The anagen hair follicles within the histoculture scalp skin produced growing hair shafts. Hair follicles could continue their cycle in histoculture; for example, apparent spontaneous catagen induction was observed both histologically and by the actual regression of the hair follicle. In addition, vellus follicles were shown to be viable at day 40 after initiation of culture. Follicle keratinocytes continued to incorporate [3H]thymidine for up to several weeks after shaft elongation had ceased. Intensive hair growth was observed in the pieces of shaved mouse skin histocultured on Gelfoam®. Isolated human and mouse hair follicles also produced growing hair shafts. By day 63 in histoculture of mouse hair follicles, the number of hair follicle-associated pluripotent (HAP) stem cells increased significantly and the follicles were intact. Gelfoam® histoculture of skin demonstrated that the hair follicle cells are the most sensitive to doxorubicin which prevented hair growth, thereby mimicking chemotherapy-induced alopecia in Gelfoam® histoculture.
J G He - One of the best experts on this subject based on the ideXlab platform.
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a randomised controlled trial comparing spontaneous healing Gelfoam patching and edge approximation plus Gelfoam patching in traumatic tympanic membrane perforation with inverted or everted edges
Clinical Otolaryngology, 2011Co-Authors: J G HeAbstract:OBJECTIVE: To compare the outcome of patients with dry traumatic tympanic membrane perforation after spontaneous healing and Gelfoam patching with or without perforation edge approximation. DESIGN: Prospective clinical study. SETTING: University-affiliated teaching hospital. PARTICIPANTS: Ninety-one patients with acute dry traumatic tympanic membrane perforation inverted or everted edges were recruited. They were randomly allocated to three groups: spontaneous healing (n=31), Gelfoam patching (n=30) and edge-approximation plus Gelfoam patching (n=30). Otoscopy and tympanometry were performed before the treatment and at follow-up visits. MAIN OUTCOME MEASURES: Healing rate, healing time, ear infection rate and morphological changes during healing process. RESULTS: The overall healing rate was 85% in the spontaneous healing group, lower than that in the two Gelfoam patching groups (97%), but the difference failed to reach a statistical significance (P>0.05). The average healing time was 30 ± 10.1 days in the spontaneous healing group, significantly longer (P<0.01) than that in the other two groups (16 ± 5.6 and 18 ± 4.7 days, respectively). Middle ear infection rate did not differ significantly (7%, 3% and 3%, respectively). Spontaneous healing resulted in formation of scabs at the perforation edges, which was effectively prevented by Gelfoam patching. CONCLUSIONS: Gelfoam patching may facilitate healing of traumatically perforated tympanic membrane. Approximation of folded perforation edges is not necessary in Gelfoam patching.
Fuminari Uehara - One of the best experts on this subject based on the ideXlab platform.
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tumor targeting salmonella typhimurium a1 r inhibits osteosarcoma angiogenesis in the in vivo Gelfoam assay visualized by color coded imaging
Anticancer Research, 2018Co-Authors: Fuminari Uehara, Yasunori Tome, Fuminori Kanaya, Tasuku Kiyuna, Takashi Murakami, Yong Zhang, Ming Zhao, Robert M HoffmanAbstract:Background We previously developed a color-coded imaging model that can quantify the length of nascent blood vessels using Gelfoam® implanted in nestin-driven green fluorescent protein (ND-GFP) nude mice. In this model, nascent blood vessels selectively express GFP. We also previously showed that osteosarcoma cells promote angiogenesis in this assay. We have also previously demonstrated the tumor-targeting bacteria Salmonella typhimurium A1-R (S. typhimurium A1-R) can inhibit or regress all tested tumor types in mouse models. The aim of the present study was to determine if S. typhimurium A1-R could inhibit osteosarcoma angiogenesis in the in vivo Gelfoam® color-coded imaging assay. Materials and methods Gelfoam® was implanted subcutaneously in ND-GFP nude mice. Skin flaps were made 7 days after implantation and 143B-RFP human osteosarcoma cells expressing red fluorescent protein (RFP) were injected into the implanted Gelfoam. After establishment of tumors in the Gelfoam®, control-group mice were treated with phosphate buffered saline via tail-vein injection (iv) and the experimental group was treated with S. typhimurium A1-R iv Skin flaps were made at day 7, 14, 21, and 28 after implantation of the Gelfoam® to allow imaging of vascularization in the Gelfoam® using a variable-magnification small-animal imaging system and confocal fluorescence microscopy. Results Nascent blood vessels expressing ND-GFP extended into the Gelfoam® over time in both groups. However, the extent of nascent blood-vessel growth was significantly inhibited by S. typhimurium A1-R treatment by day 28. Conclusion The present results indicate S. typhimurium A1-R has potential for anti-angiogenic targeted therapy of osteosarcoma.
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osteosarcoma cells enhance angiogenesis visualized by color coded imaging in the in vivo Gelfoam assay
Journal of Cellular Biochemistry, 2014Co-Authors: Shuya Yano, Fuminari Uehara, Yasunori Tome, Shinji Miwa, Yukihiko Hiroshima, Mako Yamamoto, Sumiyuki Mii, Hiroki MaeharaAbstract:We previously described a color-coded imaging model that can quantify the length of nascent blood vessels using Gelfoam® implanted in nestin-driven green fluorescent protein (ND-GFP) nude mice. In ND-GFP mice, nascent blood vessels are labeled with GFP. We report here that osteosarcoma cells promote angiogenesis in the Gelfoam® angiogenesis assay in ND-GFP mice. Gelfoam® was initially transplanted subcutaneously in the flank of transgenic ND-GFP nude mice. Seven days after transplantation of Gelfoam®, skin flaps were made and human 143B osteosarcoma cells expressing green fluorescent protein (GFP) in the nucleus and red fluorescent protein (RFP) in cytoplasm were injected into the transplanted Gelfoam®. The control-group mice had only implanted Gelfoam®. Skin flaps were made at days 14, 21, and 28 after transplantation of the Gelfoam® to allow imaging of vascularization in the Gelfoam® using a variable-magnification small animal imaging system and confocal fluorescence microscopy. ND-GFP expressing nascent blood vessels penetrated and spread into the Gelfoam® in a time-dependent manner in both control and osteosarcoma-implanted mice. ND-GFP expressing blood vessels in the Gelfoam® of the osteosarcoma-implanted mice were associated with the cancer cells and larger and longer than in the Gelfoam®-only implanted mice (P < 0.01). The results presented in this report demonstrate strong angiogenesis induction by osteosarcoma cells and suggest this process is a potential therapeutic target for this disease. J. Cell. Biochem. 115: 1490–1494, 2014. © 2014 Wiley Periodicals, Inc.
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Osteosarcoma cells enhance angiogenesis visualized by color-coded imaging in the in vivo Gelfoam® assay
Journal of Cellular Biochemistry, 2014Co-Authors: Fuminari Uehara, Shuya Yano, Yasunori Tome, Shinji Miwa, Yukihiko Hiroshima, Mako Yamamoto, Sumiyuki Mii, Hiroki Maehara, Michael Bouvet, Fuminori KanayaAbstract:We previously described a color-coded imaging model that can quantify the length of nascent blood vessels using Gelfoam® implanted in nestin-driven green fluorescent protein (ND-GFP) nude mice. In ND-GFP mice, nascent blood vessels are labeled with GFP. We report here that osteosarcoma cells promote angiogenesis in the Gelfoam® angiogenesis assay in ND-GFP mice. Gelfoam® was initially transplanted subcutaneously in the flank of transgenic ND-GFP nude mice. Seven days after transplantation of Gelfoam®, skin flaps were made and human 143B osteosarcoma cells expressing green fluorescent protein (GFP) in the nucleus and red fluorescent protein (RFP) in cytoplasm were injected into the transplanted Gelfoam®. The control-group mice had only implanted Gelfoam®. Skin flaps were made at days 14, 21, and 28 after transplantation of the Gelfoam® to allow imaging of vascularization in the Gelfoam® using a variable-magnification small animal imaging system and confocal fluorescence microscopy. ND-GFP expressing nascent blood vessels penetrated and spread into the Gelfoam® in a time-dependent manner in both control and osteosarcoma-implanted mice. ND-GFP expressing blood vessels in the Gelfoam® of the osteosarcoma-implanted mice were associated with the cancer cells and larger and longer than in the Gelfoam®-only implanted mice (P
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3‐Dimensional Tissue Is Formed From Cancer Cells In Vitro on Gelfoam®, But Not on MatrigelTM
Journal of Cellular Biochemistry, 2014Co-Authors: Yasunori Tome, Lei Zhang, Fuminari Uehara, Sumiyuki Mii, Hiroki Maehara, Michael Bouvet, Shuuya Yano, Naotoshi Sugimoto, Hiroyuki Tsuchiya, Fuminori KanayaAbstract:Cell and tissue culture can be performed on different substrates such as on plastic, in Matrigel™, and on Gelfoam®, a sponge matrix. Each of these substrates consists of a very different surface, ranging from hard and inflexible, a gel, and a sponge-matrix, respectively. Folkman and Moscona found that cell shape was tightly coupled to DNA synthesis and cell growth. Therefore, the flexibility of a substrate is important for cells to maintain their optimal shape. Human osteosarcoma cells, stably expressing a fusion protein of αv integrin and green fluorescent protein (GFP), grew as a simple monolayer without any structure formation on the surface of a plastic dish. When the osteosarcoma cells were cultured within Matrigel™, the cancer cells formed colonies but no other structures. When the cancer cells were seeded on Gelfoam®, the cells formed three-dimensional tissue-like structures. The behavior of 143B osteosarcoma cells on Gelfoam® in culture is remarkably different from those of these cells in monolayer culture or in Matrigel™. Tissue-like structures were observed only in Gelfoam® culture. The data in this report suggest a flexible structural substrate such as Gelfoam® provides a more in vivo-like culture condition than monolayer culture or MatrigelTM and that MatrigelTM does not result in actual three-dimensional culture. J. Cell. Biochem. 115: 1362–1367, 2014. © 2014 Wiley Periodicals, Inc.
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A Color-coded Imaging Model of the Interaction of αv Integrin-GFP Expressed in Osteosarcoma Cells and RFP Expressing Blood Vessels in Gelfoam® Vascularized In Vivo
Anticancer research, 2013Co-Authors: Fuminari Uehara, Shuya Yano, Yasunori Tome, Shinji Miwa, Yukihiko Hiroshima, Sumiyuki Mii, Hiroki Maehara, Michael Bouvet, Fuminori Kanaya, Robert M HoffmanAbstract:The integrin family of proteins has been shown to be involved in the malignant behavior of cells. We report here development of a color-coded imaging model that can visualize the interaction between αv integrin linked to green fluorescent protein (GFP) in osteosarcoma cells and blood vessels in Gelfoam® vascularized after implantation in red fluorescent protein (RFP) transgenic nude mice. Human 143B osteosarcoma cells expressing αv integrin-GFP were generated by transfection with an αv integrin-GFP vector. Gelfoam® (5×5 mm) was transplanted subcutaneously in transgenic RFP nude mice. The implanted Gelfoam® became highly vascularized with RFP vessels within 14 days. Skin flaps were made at days 7, 14, 21, 28 after transplantation of Gelfoam® for observing vascularization of the Gelfoam® using fluorescence imaging. Gelfoam® is a useful tool to observe angiogenesis in vivo. 143B cells (5 × 10(5)) expressing αv integrin-GFP were injected into the Gelfoam® seven days after transplantation of Gelfoam®. Seven days after cancer-cell injection, cancer cells and blood vessels were observed in the Gelfoam® by color-coded confocal microscopy via the skin flap. The 143B cells expressing αv integrin-GFP proliferated into the Gelfoam®, which contained RFP-expressing blood vessels. Strong expression of αv integrin-GFP in 143B cells was observed near RFP vessels in the Gelfoam®. The observation of the behavior of αv integrin-GFP and blood vessels will allow further understanding of the role of αv integrin in cancer cells.
Shuya Yano - One of the best experts on this subject based on the ideXlab platform.
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efficacy of a cell cycle decoying killer adenovirus on 3 d Gelfoam histoculture and tumor sphere models of chemo resistant stomach carcinomatosis visualized by fucci imaging
PLOS ONE, 2016Co-Authors: Shuya Yano, Kiyoto Takehara, Hiroshi Tazawa, Hiroyuki Kishimoto, Yasuo Urata, Shunsuke Kagawa, Toshiyoshi Fujiwara, Robert M HoffmanAbstract:Stomach cancer carcinomatosis peritonitis (SCCP) is a recalcitrant disease. The goal of the present study was to establish an in vitro-in vivo-like imageable model of SCCP to develop cell-cycle-based therapeutics of SCCP. We established 3-D Gelfoam® histoculture and tumor-sphere models of SCCP. FUCCI-expressing MKN-45 stomach cancer cells were transferred to express the fluorescence ubiquinized cell-cycle indicator (FUCCI). FUCCI-expressing MKN-45 cells formed spheres on agarose or on Gelfoam® grew into tumor-like structures with G0/G1 cancer cells in the center and S/G2 cancer cells located in the surface as indicated by FUCCI imaging when the cells fluoresced red or green, respectively. We treated FUCCI-expressing cancer cells forming SCCP tumors in Gelfoam® histoculture with OBP-301, cisplatinum (CDDP), or paclitaxel. CDDP or paclitaxel killed only cycling cancer cells and were ineffective against G1/G2 MKN-45 cells in tumors growing on Gelfoam®. In contrast, the telomerase-dependent adenovirus OBP-301 decoyed the MKN-45 cells in tumors on Gelfoam® to cycle from G0/G1 phase to S/G2 phase and reduced their viability. CDDP- or paclitaxel-treated MKN-45 tumors remained quiescent and did not change in size. In contrast, OB-301 reduced the size of the MKN-45 tumors on Gelfoam®. We examined the cell cycle-related proteins using Western blotting. CDDP increased the expression of p53 and p21 indicating cell cycle arrest. In contrast, OBP-301 decreased the expression of p53 and p21 Furthermore, OBP-301 increased the expression of E2F and pAkt as further indication of cell cycle decoy. This 3-D Gelfoam® histoculture and FUCCI imaging are powerful tools to discover effective therapy of SCCP such as OBP-301.
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Cell cycle distribution of MKN-45 cells forming tumors on Gelfoam®.
2016Co-Authors: Shuya Yano, Kiyoto Takehara, Hiroshi Tazawa, Hiroyuki Kishimoto, Yasuo Urata, Shunsuke Kagawa, Toshiyoshi Fujiwara, Robert M HoffmanAbstract:Sterile Gelfoam® sponges (Pharmacia & Upjohn, Kalamazoo, MI), prepared from porcine skin, were cut into 1 cm cubes. The Gelfoam® cubes were incubated at 37℃ in order that Gelfoam® absorbed the RPMI 1640 medium. The absorbed Gelfoam® was placed on agorose in 35 mm dishes. FUCCI-expressing MKN-45 cells were seeded on the absorbed Gelfoam® for 3D culture. MKN-45 cells formed tumor-like structures on Gelfoam® and spheres on the agarose. A. Schema of Gelfoam® cultures on agarose. B. Macroscopic appearance of MRN-45 cancer cells forming tumor-like structures on Gelfoam®. Images were acquired with the OV100 variable magnification fluorescence imager (Olympus, Japan). C. Macroscopic appearance of MKN-45 tumor-like structures growing on Gelfoam®. Images were acquired with the OV100. D. Representative images of the MKN-45 tumor-like structure on Gelfoam® using the FV1000 confocal laser scanning microscope (Olympus). E. Macroscopic images of MKN-45 spheres growing in agar. F. Microscopic FUCCI image of MKN-45 spheres.
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osteosarcoma cells enhance angiogenesis visualized by color coded imaging in the in vivo Gelfoam assay
Journal of Cellular Biochemistry, 2014Co-Authors: Shuya Yano, Fuminari Uehara, Yasunori Tome, Shinji Miwa, Yukihiko Hiroshima, Mako Yamamoto, Sumiyuki Mii, Hiroki MaeharaAbstract:We previously described a color-coded imaging model that can quantify the length of nascent blood vessels using Gelfoam® implanted in nestin-driven green fluorescent protein (ND-GFP) nude mice. In ND-GFP mice, nascent blood vessels are labeled with GFP. We report here that osteosarcoma cells promote angiogenesis in the Gelfoam® angiogenesis assay in ND-GFP mice. Gelfoam® was initially transplanted subcutaneously in the flank of transgenic ND-GFP nude mice. Seven days after transplantation of Gelfoam®, skin flaps were made and human 143B osteosarcoma cells expressing green fluorescent protein (GFP) in the nucleus and red fluorescent protein (RFP) in cytoplasm were injected into the transplanted Gelfoam®. The control-group mice had only implanted Gelfoam®. Skin flaps were made at days 14, 21, and 28 after transplantation of the Gelfoam® to allow imaging of vascularization in the Gelfoam® using a variable-magnification small animal imaging system and confocal fluorescence microscopy. ND-GFP expressing nascent blood vessels penetrated and spread into the Gelfoam® in a time-dependent manner in both control and osteosarcoma-implanted mice. ND-GFP expressing blood vessels in the Gelfoam® of the osteosarcoma-implanted mice were associated with the cancer cells and larger and longer than in the Gelfoam®-only implanted mice (P < 0.01). The results presented in this report demonstrate strong angiogenesis induction by osteosarcoma cells and suggest this process is a potential therapeutic target for this disease. J. Cell. Biochem. 115: 1490–1494, 2014. © 2014 Wiley Periodicals, Inc.
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Osteosarcoma cells enhance angiogenesis visualized by color-coded imaging in the in vivo Gelfoam® assay
Journal of Cellular Biochemistry, 2014Co-Authors: Fuminari Uehara, Shuya Yano, Yasunori Tome, Shinji Miwa, Yukihiko Hiroshima, Mako Yamamoto, Sumiyuki Mii, Hiroki Maehara, Michael Bouvet, Fuminori KanayaAbstract:We previously described a color-coded imaging model that can quantify the length of nascent blood vessels using Gelfoam® implanted in nestin-driven green fluorescent protein (ND-GFP) nude mice. In ND-GFP mice, nascent blood vessels are labeled with GFP. We report here that osteosarcoma cells promote angiogenesis in the Gelfoam® angiogenesis assay in ND-GFP mice. Gelfoam® was initially transplanted subcutaneously in the flank of transgenic ND-GFP nude mice. Seven days after transplantation of Gelfoam®, skin flaps were made and human 143B osteosarcoma cells expressing green fluorescent protein (GFP) in the nucleus and red fluorescent protein (RFP) in cytoplasm were injected into the transplanted Gelfoam®. The control-group mice had only implanted Gelfoam®. Skin flaps were made at days 14, 21, and 28 after transplantation of the Gelfoam® to allow imaging of vascularization in the Gelfoam® using a variable-magnification small animal imaging system and confocal fluorescence microscopy. ND-GFP expressing nascent blood vessels penetrated and spread into the Gelfoam® in a time-dependent manner in both control and osteosarcoma-implanted mice. ND-GFP expressing blood vessels in the Gelfoam® of the osteosarcoma-implanted mice were associated with the cancer cells and larger and longer than in the Gelfoam®-only implanted mice (P
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abstract 702 chinese medicine formulation lq inhibits fucci expressing hela cells at the g0 g1phase in 2d and 3d culture and invasion with Gelfoam 3d culture
Cancer Research, 2013Co-Authors: Lei Zhang, Shuya Yano, Robert M HoffmanAbstract:Proceedings: AACR 104th Annual Meeting 2013; Apr 6-10, 2013; Washington, DC Traditional Chinese Medicine formula LQ has previously demonstrated anti-tumor and anti-metastasis efficacy. Here, we used fluorescence ubiquitination-based cell cycle indicator (FUCCI) to monitor cell cycle arrest after LQ treatment. FUCCI-HeLa cells were cultured in two dimensional (2D) monolayer, 3D Matrigel®, and 3D Gelform®. Changes of cell cycle status after 24 hours LQ treatment (90mg/ml) were observed using the Olympus FV1000 confocal imaging system. Paclitaxel (Taxol) was used as the positive control which induced a G2/M block of cell cycle. LQ blocked FUCCI-HeLa cells in the G/G1 phase of the cell cycle in all cultures. In monolayer culture, the control had approximately 45% of the cells in G2/M phase. In contrast, the LQ-treated cells were mostly in the G/G1 phase (>90%). In 3D dimensional culture (Matrigel®), cancer cells formed spheres. The colonies in control group had 40% of the cells in G2/M phase, but only 15% in LQ-treated culture. The cancer cells at the surface of the sphere were mostly in G2/M phase, and the cancer cells in the center of colonies were in G/G1 phase. In 3D Gelform® culture, cells grew along the structures of the Gelform®. The control culture had approximately 45% of cells in G2/M phase. In contrast, the cells in LQ-treated culture were mostly in G/G1 phase (>80%). The cells in control group migrated to 250∼300 μm deep in the Gelfoam®, but only 150∼200 μm deep in LQ-treated culture. FUCCI-HeLa cells in different culture systems demonstrated the efficacy of LQ on the cell cycle and migration. Citation Format: Lei Zhang, Chengyu Wu, Shuya Yano, Robert M. Hoffman. Chinese Medicine formulation LQ inhibits FUCCI-expressing HeLa cells at the G/G1 phase in 2D and 3D culture and invasion with Gelfoam® 3D culture. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 702. doi:10.1158/1538-7445.AM2013-702
Yasunori Tome - One of the best experts on this subject based on the ideXlab platform.
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tumor targeting salmonella typhimurium a1 r inhibits osteosarcoma angiogenesis in the in vivo Gelfoam assay visualized by color coded imaging
Anticancer Research, 2018Co-Authors: Fuminari Uehara, Yasunori Tome, Fuminori Kanaya, Tasuku Kiyuna, Takashi Murakami, Yong Zhang, Ming Zhao, Robert M HoffmanAbstract:Background We previously developed a color-coded imaging model that can quantify the length of nascent blood vessels using Gelfoam® implanted in nestin-driven green fluorescent protein (ND-GFP) nude mice. In this model, nascent blood vessels selectively express GFP. We also previously showed that osteosarcoma cells promote angiogenesis in this assay. We have also previously demonstrated the tumor-targeting bacteria Salmonella typhimurium A1-R (S. typhimurium A1-R) can inhibit or regress all tested tumor types in mouse models. The aim of the present study was to determine if S. typhimurium A1-R could inhibit osteosarcoma angiogenesis in the in vivo Gelfoam® color-coded imaging assay. Materials and methods Gelfoam® was implanted subcutaneously in ND-GFP nude mice. Skin flaps were made 7 days after implantation and 143B-RFP human osteosarcoma cells expressing red fluorescent protein (RFP) were injected into the implanted Gelfoam. After establishment of tumors in the Gelfoam®, control-group mice were treated with phosphate buffered saline via tail-vein injection (iv) and the experimental group was treated with S. typhimurium A1-R iv Skin flaps were made at day 7, 14, 21, and 28 after implantation of the Gelfoam® to allow imaging of vascularization in the Gelfoam® using a variable-magnification small-animal imaging system and confocal fluorescence microscopy. Results Nascent blood vessels expressing ND-GFP extended into the Gelfoam® over time in both groups. However, the extent of nascent blood-vessel growth was significantly inhibited by S. typhimurium A1-R treatment by day 28. Conclusion The present results indicate S. typhimurium A1-R has potential for anti-angiogenic targeted therapy of osteosarcoma.
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osteosarcoma cells enhance angiogenesis visualized by color coded imaging in the in vivo Gelfoam assay
Journal of Cellular Biochemistry, 2014Co-Authors: Shuya Yano, Fuminari Uehara, Yasunori Tome, Shinji Miwa, Yukihiko Hiroshima, Mako Yamamoto, Sumiyuki Mii, Hiroki MaeharaAbstract:We previously described a color-coded imaging model that can quantify the length of nascent blood vessels using Gelfoam® implanted in nestin-driven green fluorescent protein (ND-GFP) nude mice. In ND-GFP mice, nascent blood vessels are labeled with GFP. We report here that osteosarcoma cells promote angiogenesis in the Gelfoam® angiogenesis assay in ND-GFP mice. Gelfoam® was initially transplanted subcutaneously in the flank of transgenic ND-GFP nude mice. Seven days after transplantation of Gelfoam®, skin flaps were made and human 143B osteosarcoma cells expressing green fluorescent protein (GFP) in the nucleus and red fluorescent protein (RFP) in cytoplasm were injected into the transplanted Gelfoam®. The control-group mice had only implanted Gelfoam®. Skin flaps were made at days 14, 21, and 28 after transplantation of the Gelfoam® to allow imaging of vascularization in the Gelfoam® using a variable-magnification small animal imaging system and confocal fluorescence microscopy. ND-GFP expressing nascent blood vessels penetrated and spread into the Gelfoam® in a time-dependent manner in both control and osteosarcoma-implanted mice. ND-GFP expressing blood vessels in the Gelfoam® of the osteosarcoma-implanted mice were associated with the cancer cells and larger and longer than in the Gelfoam®-only implanted mice (P < 0.01). The results presented in this report demonstrate strong angiogenesis induction by osteosarcoma cells and suggest this process is a potential therapeutic target for this disease. J. Cell. Biochem. 115: 1490–1494, 2014. © 2014 Wiley Periodicals, Inc.
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Osteosarcoma cells enhance angiogenesis visualized by color-coded imaging in the in vivo Gelfoam® assay
Journal of Cellular Biochemistry, 2014Co-Authors: Fuminari Uehara, Shuya Yano, Yasunori Tome, Shinji Miwa, Yukihiko Hiroshima, Mako Yamamoto, Sumiyuki Mii, Hiroki Maehara, Michael Bouvet, Fuminori KanayaAbstract:We previously described a color-coded imaging model that can quantify the length of nascent blood vessels using Gelfoam® implanted in nestin-driven green fluorescent protein (ND-GFP) nude mice. In ND-GFP mice, nascent blood vessels are labeled with GFP. We report here that osteosarcoma cells promote angiogenesis in the Gelfoam® angiogenesis assay in ND-GFP mice. Gelfoam® was initially transplanted subcutaneously in the flank of transgenic ND-GFP nude mice. Seven days after transplantation of Gelfoam®, skin flaps were made and human 143B osteosarcoma cells expressing green fluorescent protein (GFP) in the nucleus and red fluorescent protein (RFP) in cytoplasm were injected into the transplanted Gelfoam®. The control-group mice had only implanted Gelfoam®. Skin flaps were made at days 14, 21, and 28 after transplantation of the Gelfoam® to allow imaging of vascularization in the Gelfoam® using a variable-magnification small animal imaging system and confocal fluorescence microscopy. ND-GFP expressing nascent blood vessels penetrated and spread into the Gelfoam® in a time-dependent manner in both control and osteosarcoma-implanted mice. ND-GFP expressing blood vessels in the Gelfoam® of the osteosarcoma-implanted mice were associated with the cancer cells and larger and longer than in the Gelfoam®-only implanted mice (P
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3‐Dimensional Tissue Is Formed From Cancer Cells In Vitro on Gelfoam®, But Not on MatrigelTM
Journal of Cellular Biochemistry, 2014Co-Authors: Yasunori Tome, Lei Zhang, Fuminari Uehara, Sumiyuki Mii, Hiroki Maehara, Michael Bouvet, Shuuya Yano, Naotoshi Sugimoto, Hiroyuki Tsuchiya, Fuminori KanayaAbstract:Cell and tissue culture can be performed on different substrates such as on plastic, in Matrigel™, and on Gelfoam®, a sponge matrix. Each of these substrates consists of a very different surface, ranging from hard and inflexible, a gel, and a sponge-matrix, respectively. Folkman and Moscona found that cell shape was tightly coupled to DNA synthesis and cell growth. Therefore, the flexibility of a substrate is important for cells to maintain their optimal shape. Human osteosarcoma cells, stably expressing a fusion protein of αv integrin and green fluorescent protein (GFP), grew as a simple monolayer without any structure formation on the surface of a plastic dish. When the osteosarcoma cells were cultured within Matrigel™, the cancer cells formed colonies but no other structures. When the cancer cells were seeded on Gelfoam®, the cells formed three-dimensional tissue-like structures. The behavior of 143B osteosarcoma cells on Gelfoam® in culture is remarkably different from those of these cells in monolayer culture or in Matrigel™. Tissue-like structures were observed only in Gelfoam® culture. The data in this report suggest a flexible structural substrate such as Gelfoam® provides a more in vivo-like culture condition than monolayer culture or MatrigelTM and that MatrigelTM does not result in actual three-dimensional culture. J. Cell. Biochem. 115: 1362–1367, 2014. © 2014 Wiley Periodicals, Inc.
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A Color-coded Imaging Model of the Interaction of αv Integrin-GFP Expressed in Osteosarcoma Cells and RFP Expressing Blood Vessels in Gelfoam® Vascularized In Vivo
Anticancer research, 2013Co-Authors: Fuminari Uehara, Shuya Yano, Yasunori Tome, Shinji Miwa, Yukihiko Hiroshima, Sumiyuki Mii, Hiroki Maehara, Michael Bouvet, Fuminori Kanaya, Robert M HoffmanAbstract:The integrin family of proteins has been shown to be involved in the malignant behavior of cells. We report here development of a color-coded imaging model that can visualize the interaction between αv integrin linked to green fluorescent protein (GFP) in osteosarcoma cells and blood vessels in Gelfoam® vascularized after implantation in red fluorescent protein (RFP) transgenic nude mice. Human 143B osteosarcoma cells expressing αv integrin-GFP were generated by transfection with an αv integrin-GFP vector. Gelfoam® (5×5 mm) was transplanted subcutaneously in transgenic RFP nude mice. The implanted Gelfoam® became highly vascularized with RFP vessels within 14 days. Skin flaps were made at days 7, 14, 21, 28 after transplantation of Gelfoam® for observing vascularization of the Gelfoam® using fluorescence imaging. Gelfoam® is a useful tool to observe angiogenesis in vivo. 143B cells (5 × 10(5)) expressing αv integrin-GFP were injected into the Gelfoam® seven days after transplantation of Gelfoam®. Seven days after cancer-cell injection, cancer cells and blood vessels were observed in the Gelfoam® by color-coded confocal microscopy via the skin flap. The 143B cells expressing αv integrin-GFP proliferated into the Gelfoam®, which contained RFP-expressing blood vessels. Strong expression of αv integrin-GFP in 143B cells was observed near RFP vessels in the Gelfoam®. The observation of the behavior of αv integrin-GFP and blood vessels will allow further understanding of the role of αv integrin in cancer cells.
