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Shuji Horibe - One of the best experts on this subject based on the ideXlab platform.
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autologous bone marrow Stromal Cell transplantation for repair of full thickness articular cartilage defects in human patellae two case reports
Cell Transplantation, 2004Co-Authors: Shigeyuki Wakitani, Tomoki Mitsuoka, Norimasa Nakamura, Yukiyoshi Toritsuka, Yukio Nakamura, Shuji HoribeAbstract:This study assessed the effectiveness of autologous bone marrow Stromal Cell transplantation for the repair of full-thickness articular cartilage defects in the patellae of a 26-year-old female and...
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autologous bone marrow Stromal Cell transplantation for repair of full thickness articular cartilage defects in human patellae two case reports
Cell Transplantation, 2004Co-Authors: Shigeyuki Wakitani, Tomoki Mitsuoka, Norimasa Nakamura, Yukiyoshi Toritsuka, Yukio Nakamura, Shuji HoribeAbstract:This study assessed the effectiveness of autologous bone marrow Stromal Cell transplantation for the repair of full-thickness articular cartilage defects in the patellae of a 26-year-old female and a 44-year-old male. These two patients presented in our clinic because their knee pain prevented them from walking normally. After thorough examination, we concluded that the knee pain was due to the injured articular cartilage and decided to repair the defect with bone marrow Stromal Cell transplantation. Three weeks before transplantation, bone marrow was aspirated from the iliac crest of each patient. After erythrocytes had been removed by use of dextran, the remaining nucleated Cells were placed in culture. When the attached Cells reached subconfluence, they were passaged to expand in culture. Adherent Cells were subsequently collected, embedded in a collagen gel, transplanted into the articular cartilage defect in the patellae, and covered with autologous periosteum. Six months after transplantation, clinical symptoms (pain and walking ability) had improved significantly and the improvement has remained in effect (5 years and 9 months posttransplantation in one case, and 4 years in the other), and both patients have been satisfied with the outcome. As early as 2 months after transplantation, the defects were covered with tissue that showed slight metachromatic staining. Two years after the first and 1 year after the second transplantation, arthroscopy was performed and the defects were repaired with fibrocartilage. Results indicate autologous bone marrow Stromal Cell transplantation is an effective approach in promoting the repair of articular cartilage defects.
Joel S Greenberger - One of the best experts on this subject based on the ideXlab platform.
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radioresistance of serpinb3a mice and derived hematopoietic and marrow Stromal Cell lines
Radiation Research, 2019Co-Authors: Stephanie Thermozier, Michael W Epperly, Wen Hou, Xichen Zhang, Renee Fisher, Bing Liu, Ivet Bahar, Hong Wang, Joel S GreenbergerAbstract:Serpins are a group of serine-proteases involved in multiple signal transduction pathways in mammalian Cells. In particular, Serpinb3a is involved in the lysosomal necrosis Cell death pathway with components that overlap with radiation-induced apoptosis. We investigated the radiation response of Serpinb3a-/- mice compared to Serpinb3a+/+ mice on the Balb/c background. Serpinb3a-/- mice showed significant radioresistance to a dose of 8.0 Gy total-body irradiation, compared to Serpinb3a+/+ Balb/c mice. Long-term bone marrow cultures from Serpinb3a-/- mice showed increased longevity. In clonogenic survival assays, fresh bone marrow hematopoietic progenitors, as well as clonal interleukin-3 (IL-3)-dependent hematopoietic progenitor and bone marrow Stromal Cell lines from Serpinb3a-/- mice were radioresistant. Serpinb3a-/- mouse bone marrow-derived Stromal Cell lines had increased baseline and postirradiation antioxidant capacity. Serpinb3a-/- bone marrow Stromal Cells showed increased radiation-induced RNA transcripts for MnSOD and p21, and decreased levels of p53 and TGF-b. Both irradiated Serpinb3a-/- mouse bone marrow Stromal Cell lines and plasma removed from total-body irradiated mice had decreased levels of expression of stress response and inflammation-associated proteins. Abrogation of Serpinb3a may be a potential new target for mitigation of radiation effects.
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radiation resistance of c57bl 6 mouse bone marrow Stromal Cell lines induced by apoptosis inhibitor jp4 039 or necroptosis inhibitor necrostatin 1 but not by simultaneous administration of both mitigators
Blood, 2017Co-Authors: Zhiyang Wei, Michael W Epperly, Donna Shields, Wen Hou, Darcy Franicola, Xichen Zhang, Renee Fisher, Hulya Bayir, Peter Wipf, Joel S GreenbergerAbstract:Abstract Introduction: Necroptosis is a TNFa activated, caspase-independent Cell death pathway that involves active disintegration of mitochondrial, lysosomal and plasma membranes (Vandenabeele, et al, Nat Rev Mol Cell Biol., 2010 Oct; 11(10):700-14). We measured clonogenic radiation survival of C57/Bl6 mouse bone marrow Stromal Cell lines in the presence of a mitochondrial targeted inhibitor of apoptosis (JP4-039), an inhibitor of necroptosis (Necrostatin-1) or both drugs. Materials and Methods: C57BL/6 mouse long term bone marrow cultures (LTBMCs) were established from adult female C57Bl/6 mice. Bone marrow Stromal Cell lines were derived from the adherent layers of LTBMCs. Radiosensitivity of C57Bl/6 bone marrow Stromal Cell lines and fresh bone marrow CFU-GEMM (semisolid medium with added hemopoietic growth factors) was measured in clonogenic radiation survival curves. Stromal Cells were irradiated to doses of 0 to 8 Gy, plated in T-75 tissue culture flasks, incubated for 8 days at 37oC, stained with crystal violet and colonies of greater than 50 Cells counted at day 7. CFU-GEMMs from fresh bone marrow were scored at day 14. Both cultured Cell populations were tested with addition of radiation mitigators: JP4-039 (10µM), Necrostatin-1 (40µM), or both drugs added to culture medium 30 minutes post irradiation. C57BL/6 mouse LTBMCs generated hemopoietic Cells for 20 weeks. Marrow Stromal Cell lines were derived from 20 week old hemopoieticly-inactive culture adherent layer Cells. Statistical analysis was performed using Student's t-test, with a two-tail p-value Results: Radiation survival curves showed that C57Bl/6 fresh bone marrow CFU-GEMM and Stromal Cell lines were comparable to those for other mouse strains (CFU-GEMM Do = 1.67 ± 0.02, Stromal Cells Do=1.43 ± 0.14). Both fresh marrow CFU-GEMM and Stromal Cells (ň = 5.26 ± 0.72) were radioresistant when grown in presence of JP4-039 (ň = 8.85 ± 1.23, p = 0.012) or necrostatin-1 (n=7.97 ± 1.12, p = 0.024). Addition of both JP4-039 and Necrostatin-1 (ň = 5.79 ± 1.44) at the same time after irradiation (30 min) did not improve clonogenic survival of either fresh marrow CFU-GEMM or Stromal Cell lines. Conclusions: Both freshly explanted CFU-GEMM marrow hematopoietic progenitor Cells and LTBMC derived marrow Stromal Cell lines display radioresistance when treated with each of two radiation mitigators that target different Cell death pathways. While each drug delivered independently induced radioresistance, combined delivery of JP4-039 and Necrostatin-1 at 30 min after irradiation did not. Thus, the timing of delivery of drugs that target different Cell death pathways is likely critical in designing effective protocols for radiation mitigation. Disclosures No relevant conflicts of interest to declare.
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transformed phenotype of bone marrow Stromal Cell lines derived from k14e7 fancd2 mice
Blood, 2015Co-Authors: Aranee Sivanathan, Michael W Epperly, Donna Shields, Darcy Franicola, Xichen Zhang, Shaonan Cao, Joel S GreenbergerAbstract:To determine whether cytokeratin 14 promoter linked expression of the Human Papilloma Virus (HPV) oncogene detectably influenced biologic parameters of Cell phenotypes other than squamous epithelium, continuous bone marrow cultures were derived from K14E7 Fancd2 -/- mice (Park, et al., Cancer Research, 70(23): 9959-9968, 2010). Long-term bone marrow cultures derived from K14E7 Fancd2 -/- , control Fancd2 -/- (129/Sv), K14E7 (FVB/N), and wild type 129/Sv X FVB/N F1 mice were evaluated for longevity of hematopoiesis in long-term cultures and Stromal Cell lines were derived from each. Similar to Fancd2 -/- mouse, long-term marrow cultures, K14E7 Fancd2 -/- marrow cultures demonstrated decreased longevity of hematopoiesis with cessation of production of multi-lineage colony forming progenitor Cells after 14 weeks. In contrast, wild type F1 and K14E7 long-term marrow cultures continued to produce hematopoietic Cells for a significantly longer duration 25 weeks (p=0.0257). Bone marrow Stromal and IL-3 dependent hematopoietic Cell lines were derived from each genotype marrow culture. K14E7 Fancd2 -/- hematopoietic Cells showed reversal of the radiation resistance of Fancd2 -/- IL-3 dependent Cell lines (D 0 of 1.34 ± 0.197Gy, n 4.0 ± 0.9 compared to D 0 2.213 ± 0.124 Gy (p = 0.0284), n 3.3 ± 0.8. Thus, one phenotypic difference associated with K14E7 oncogene expression was reversal of radioresistance of Fancd2 -/- hematopoietic Cells. In contrast, bone marrow Stromal Cell lines from K14E7 Fancd2 -/- remained radiosensitive similar to those from Fancd2 -/- mice. K14E7 and wild type F1 marrow Stromal Cell lines showed intermediate radioresistance (p= 0.1759). To determine whether the E6/E7 oncogene had a biological effect in tissues other than squamous epithelium, tissue analysis for cytokeratins 13, 14, 6, and 10 was carried out. Cytokeratin 14 was detected only in squamous Cells of the esophagus and oral cavity, not in bone marrow. E6/E7 oncogene was detected only in squamous Cell lines expressing cytokeratin 14. However, bone marrow Stromal Cell lines from K14E7 Fancd2 -/- marrow cultures demonstrated a unique phenomenon of Cellular density, piling up and formation of tumors in vitro . Each of 22 single Cell derived clonal sub-lines of K14E7 Fancd2 -/- Stromal Cell lines demonstrated the same transformed phenotype. These data provide support for indirect effects of the E7 oncogene linked to the K14 promoter in Fancd2 -/- hematopoietic and mesenchymal stem Cell tissues. Supported by research grant NIAID/NIH, U19A168021. Disclosures No relevant conflicts of interest to declare.
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quantitation of murine hematopoietic stem Cells in vitro by limiting dilution analysis of cobblestone area formation on a clonal Stromal Cell line
Experimental Hematology, 1993Co-Authors: S Neben, P Anklesaria, Joel S Greenberger, Peter MauchAbstract:Murine hematopoietic stem Cells with varying proliferative capacity can be assayed by limiting dilution analysis of "cobblestone area" (CA) formation on Stromal layers in microlong-term bone marrow cultures. Cobblestone area forming Cell (CAFC) frequency determined at early time points (day 7) correlates with mature stem Cells measured as day 8 CFU-S, whereas CAFC frequency determined at day 28 correlates more closely with long-term marrow repopulating ability. Here we report a modification of the CAFC assay in which a clonal bone marrow Stromal Cell line, GB1/6, is substituted for fresh marrow-derived Stromal layers. This modification simplifies the initial culture setup, eliminates inhomogeneities in the Stromal layer and reduces the need for mice. Normal bone marrow CAFC frequencies were the same for both types of Stromal Cell underlayer, demonstrating the ability of a clonal Cell line to completely replace the heterogeneous microenvironment of fresh stroma for in vitro stem Cell support.
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adipogenesis in a myeloid supporting bone marrow Stromal Cell line
Journal of Cellular Biochemistry, 1992Co-Authors: Jeffery M Gimble, Kellee Youkhana, Xianxin Hua, Kay L Medina, Mary Sullivan, Helen B Bass, Chisun S Wang, Joel S GreenbergerAbstract:The bone marrow stroma contains pre-adipocyte Cells which are part of the hemopoietic microenvironment. Cloned Stromal Cell lines differ both in their ability to support myeloid and lymphoid development and in their ability to undergo adipocyte differentiation in vitro. These processes have been examined in the +/+2.4 murine Stromal Cell line and compared to other Stromal and pre-adipocyte Cell lines. In long-term cultures, the +/+2.4 Stromal Cells support myeloid Cell growth, consistent with their expression of macrophage-colony stimulating factor mRNA. However, despite the presence of mRNA for the lymphoid supportive cytokines interleukins 6 and 7, +/+2.4 Cells failed to support Stromal Cell dependent B lineage lymphoid Cells in vitro, suggesting that these Stromal Cells exhibit only a myelopoietic support function. The +/+2.4 Cells differentiate into adipocytes spontaneously when cultured in 10% fetal bovine serum. The process of adipogenesis can be accelerated by a number of agonists based on morphologic and gene marker criteria. Following induction with hydrocortisone, methylisobutylxanthine, indomethacin, and insulin in combination, a time dependent increase in the steady state mRNA and enzyme activity levels of the following adipocyte specific genes was observed: adipocyte P2, adipsin, CAAT/enhancer binding protein, and lipoprotein lipase. In contrast, adipogenesis was accompanied by a slight decrease in the signal intensity of the macrophage-colony stimulating factor mRNA level, similar to that which has been reported in other bone marrow Stromal Cell lines. These data demonstrate that although the lympho-hematopoietic support function of pre-adipocyte bone marrow Stromal Cell lines is heterogeneous, they share a common mechanism of adipogenesis.
Krampera Mauro - One of the best experts on this subject based on the ideXlab platform.
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Emerging data supporting Stromal Cell therapeutic potential in cancer: reprogramming Stromal Cells of the tumor microenvironment for anti-cancer effects
'Cancer Biology and Medicine', 2020Co-Authors: Kamdje, Armel H Nwabo, Etet, Paul F Seke, Simo, Richard Tagne, Vecchio Lorella, Lukong, Kiven Erique, Krampera MauroAbstract:After more than a decade of controversy on the role of Stromal Cells in the tumor microenvironment, the emerging data shed light on pro-tumorigenic and potential anti-cancer factors, as well as on the roots of the discrepancies. We discuss the pro-tumorigenic effects of Stromal Cells, considering the effects of tumor drivers like hypoxia and tumor stiffness on these Cells, as well as Stromal Cell-mediated adiposity and immunosuppression in the tumor microenvironment, and cancer initiating Cells' Cellular senescence and adaptive metabolism. We summarize the emerging data supporting Stromal Cell therapeutic potential in cancer, discuss the possibility to reprogram Stromal Cells of the tumor microenvironment for anti-cancer effects, and explore some causes of discrepancies on the roles of Stromal Cells in cancer in the available literature
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HS-5 and HS-27A Stromal Cell Lines to Study Bone Marrow Mesenchymal Stromal Cell-Mediated Support to Cancer Development
'Frontiers Media SA', 2020Co-Authors: Adamo Annalisa, Delfino Pietro, Gatti Alessandro, Bonato Alice, Takam Kamga Paul, Bazzoni Riccardo, Ugel Stefano, Mercuri Angela, Caligola Simone, Krampera MauroAbstract:In this study, we compared the overall gene and pathway expression profiles of HS-5 and HS-27A Stromal Cell lines with those of primary bone marrow MSCs to verify if they can be considered a reliable alternative tool for evaluating the contribution of MSCs in tumor development and immunomodulation. Indeed, due to their easier manipulation in vitro as compared to primary MSC cultures, several published studies took advantage of Stromal Cell lines to assess the biological mechanisms mediated by Stromal Cells in influencing tumor biology and immune responses. However, the process carried out to obtain immortalized Cell lines could profoundly alter gene expression profile, and consequently their biological characteristics, leading to debatable results. Here, we evaluated the still undisclosed similarities and differences between HS-5, HS-27A Cell lines and primary bone marrow MSCs in the context of tumor development and immunomodulation. Furthermore, we assessed by standardized immunological assays the capability of the Cell lines to reproduce the general mechanisms of MSC immunoregulation. We found that only HS-5 Cell line could be suitable to reproduce not only the MSC capacity to influence tumor biology, but also to evaluate the molecular mechanisms underlying tumor immune escape mediated by stroma Cells. However, HS-5 pre-treatment with inflammatory cytokines, that normally enhances the immunosuppressive activity of primary MSCs, did not reproduce the same MSCs behavior, highlighting the necessity to accurately set up in vitro assays when HS-5 Cell line is used instead of its primary counterpart
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HS-5 and HS-27A Stromal Cell Lines to Study Bone Marrow Mesenchymal Stromal Cell-Mediated Support to Cancer Development
'Frontiers Media SA', 2020Co-Authors: Adamo Annalisa, Delfino Pietro, Gatti Alessandro, Bonato Alice, Takam Kamga Paul, Bazzoni Riccardo, Ugel Stefano, Mercuri Angela, Caligola Simone, Krampera MauroAbstract:International audienceIn this study, we compared the overall gene and pathway expression profiles of HS-5 and HS-27A Stromal Cell lines with those of primary bone marrow MSCs to verify if they can be considered a reliable alternative tool for evaluating the contribution of MSCs in tumor development and immunomodulation. Indeed, due to their easier manipulation in vitro as compared to primary MSC cultures, several published studies took advantage of Stromal Cell lines to assess the biological mechanisms mediated by Stromal Cells in influencing tumor biology and immune responses. However, the process carried out to obtain immortalized Cell lines could profoundly alter gene expression profile, and consequently their biological characteristics, leading to debatable results. Here, we evaluated the still undisclosed similarities and differences between HS-5, HS-27A Cell lines and primary bone marrow MSCs in the context of tumor development and immunomodulation. Furthermore, we assessed by standardized immunological assays the capability of the Cell lines to reproduce the general mechanisms of MSC immunoregulation. We found that only HS-5 Cell line could be suitable to reproduce not only the MSC capacity to influence tumor biology, but also to evaluate the molecular mechanisms underlying tumor immune escape mediated by stroma Cells. However, HS-5 pre-treatment with inflammatory cytokines, that normally enhances the immunosuppressive activity of primary MSCs, did not reproduce the same MSCs behavior, highlighting the necessity to accurately set up in vitro assays when HS-5 Cell line is used instead of its primary counterpart
Shigeyuki Wakitani - One of the best experts on this subject based on the ideXlab platform.
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autologous bone marrow Stromal Cell transplantation for repair of full thickness articular cartilage defects in human patellae two case reports
Cell Transplantation, 2004Co-Authors: Shigeyuki Wakitani, Tomoki Mitsuoka, Norimasa Nakamura, Yukiyoshi Toritsuka, Yukio Nakamura, Shuji HoribeAbstract:This study assessed the effectiveness of autologous bone marrow Stromal Cell transplantation for the repair of full-thickness articular cartilage defects in the patellae of a 26-year-old female and...
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autologous bone marrow Stromal Cell transplantation for repair of full thickness articular cartilage defects in human patellae two case reports
Cell Transplantation, 2004Co-Authors: Shigeyuki Wakitani, Tomoki Mitsuoka, Norimasa Nakamura, Yukiyoshi Toritsuka, Yukio Nakamura, Shuji HoribeAbstract:This study assessed the effectiveness of autologous bone marrow Stromal Cell transplantation for the repair of full-thickness articular cartilage defects in the patellae of a 26-year-old female and a 44-year-old male. These two patients presented in our clinic because their knee pain prevented them from walking normally. After thorough examination, we concluded that the knee pain was due to the injured articular cartilage and decided to repair the defect with bone marrow Stromal Cell transplantation. Three weeks before transplantation, bone marrow was aspirated from the iliac crest of each patient. After erythrocytes had been removed by use of dextran, the remaining nucleated Cells were placed in culture. When the attached Cells reached subconfluence, they were passaged to expand in culture. Adherent Cells were subsequently collected, embedded in a collagen gel, transplanted into the articular cartilage defect in the patellae, and covered with autologous periosteum. Six months after transplantation, clinical symptoms (pain and walking ability) had improved significantly and the improvement has remained in effect (5 years and 9 months posttransplantation in one case, and 4 years in the other), and both patients have been satisfied with the outcome. As early as 2 months after transplantation, the defects were covered with tissue that showed slight metachromatic staining. Two years after the first and 1 year after the second transplantation, arthroscopy was performed and the defects were repaired with fibrocartilage. Results indicate autologous bone marrow Stromal Cell transplantation is an effective approach in promoting the repair of articular cartilage defects.
Saul L Offman - One of the best experts on this subject based on the ideXlab platform.
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signet ring Stromal Cell tumor a legitimate benign mimic of krukenberg tumor
Archives of Pathology & Laboratory Medicine, 2018Co-Authors: Ryan Decoste, Saul L OffmanAbstract:Signet ring Stromal Cell tumor is a rare, benign ovarian neoplasm thought to arise from ovarian Stromal Cells. The pathophysiology of these tumors is poorly understood. They present in women in a wide age range, often with nonspecific symptoms including lower abdominal or pelvic pain. Their morphologic appearance raises a critical differential diagnosis of Krukenberg tumor, an aggressive malignancy with significant implications for patient management. For this reason, it is important for the pathologist to be aware of signet ring Stromal Cell tumor and its differentiating features, including useful histochemical and immunohistochemical ancillary tests. These tumors are curable with surgical excision, and there have been no recurrences or metastases among reported cases.