cd44 positive selection  (R&D Systems)

Journal: Current Issues in Molecular Biology

Article Title: Oncological Properties of Intravenous Leiomyomatosis: Involvement of Mesenchymal Tumor Stem-Like Cells

doi: 10.3390/cimb43020084

Figure Lengend Snippet: Differential expressions of PSMB9/β1i, CALPONIN h1 and CD44 in human uterine mesenchymal tumors and uterine LANT-like tumor.

Article Snippet: We isolated a candidate population of CD44-positive SK-LMS-1 subclone as human Ut-LMS stem-like cells from human Ut-LMS primary cells and SK-LMS-1 cells, using the CD44-positive selection method with MagCellect Plus Human CD44+ Cell Isolation Kit (R&D Systems, Inc., Minneapolis, MN, USA).

Techniques: Expressing

Journal: Current Issues in Molecular Biology

Article Title: Oncological Properties of Intravenous Leiomyomatosis: Involvement of Mesenchymal Tumor Stem-Like Cells

doi: 10.3390/cimb43020084

Figure Lengend Snippet: Differential expressions of factors cyclin B, cyclin E, caveolin 1, Ki-67, LMP2/b1i, and CD44 as potential biomarkers in intravenous leiomyomatosis and intravenous leiomyosarcoma. ( A ) Extensive intravascular growth of focal cellular smooth muscle is noted, forming worm-like plugs within veins. Focal hydropic change and components of endometrial glands are also observed. CD44-positive cells in the external tissue of intravenous are shown in the box a . CD44-positive cells in the internal tissue of intravenous are shown in the box b . ( B ) Intravenous leiomyosarcoma is shown in the upper right photograph. In uterine leiomyosarcoma, spindle-shaped tumor cells with round nuclei proliferate solidly. Compared with leiomyoma, uterine leiomyosarcoma shows increased nuclear density, nuclear hypertrophy, nuclear irregularities, and increased fission. ( C ) Immunohistochemistry of intravenous leiomyomatosis and intravenous leiomyosarcoma tissue sections was performed using appropriate monoclonal antibodies with standard procedures. The five tissue sites were randomly selected from intravenous leiomyomatosis’s internal and external tissues (i.e., normal uterine leiomyoma). As with uterine leiomyoma, the five tissue sites were randomly selected from the internal tissue of intravenous leiomyosarcoma. In a 40× field of view, the positive rates of the six factors were calculated in the five tissue sites. The positive rates at the sites of each tissue are shown in the scatter plot. Int.UL: Intravenous leiomyomatosis, Int.uLMS: Intravenous uterine leiomyosarcoma.

Article Snippet: We isolated a candidate population of CD44-positive SK-LMS-1 subclone as human Ut-LMS stem-like cells from human Ut-LMS primary cells and SK-LMS-1 cells, using the CD44-positive selection method with MagCellect Plus Human CD44+ Cell Isolation Kit (R&D Systems, Inc., Minneapolis, MN, USA).

Techniques: Immunohistochemistry

Journal: Current Issues in Molecular Biology

Article Title: Oncological Properties of Intravenous Leiomyomatosis: Involvement of Mesenchymal Tumor Stem-Like Cells

doi: 10.3390/cimb43020084

Figure Lengend Snippet: Induction of micrometastases in alveolar tissues derived from BALB/c nu/nu mice xenografted with human CD44-positive SK-LMS-1 stem-like cells. ( A ) Photographs show xenografts derived from BALB/c nu/nu mice inoculated with CD44-negative SK-LMS-1 cells or CD44-positive SK-LMS-1 cells at 2 months after the injection. Xenografts derived from BALB/c nu/nu mice inoculated with CD44-positive SK-LMS-1 cells were highly angiogenic. Data are representative of one of ten independent experiments obtained with different xenograft samples derived from CD44-negative SK-LMS-1 cells or CD44-positive SK-LMS-1 cells. The photographs show the pathology in metastasizes of respiratory tissue derived from the primary tumor on the left side of the second mammary fat pad, and micrometastases in the alveolar tissues of BALB/c nu/nu mice xenografted with CD44-negative SK-LMS-1 cells or CD44-positive SK-LMS-1 cells at 2 months after injection. In xenograft experiments using BALB/c nu/nu mice, the tumorigenicity of CD44-negative SK-LMS-1 cell and CD44-positive SK-LMS-1 cell xenografts was similar. Data are representative of one of ten independent experiments obtained with different xenograft samples derived from CD44-negative SK-LMS-1 cells or CD44-positive SK-LMS-1 cells. Metastatic tissues are indicated by black arrows. Micrometastasis in the alveoli are indicated by red arrows. ( B ) CD44-negative SK-LMS-1 cells or CD44-positive SK-LMS-1 cells were harvested from the cell culture using trypsin, washed, and resuspended in PBS (2 × 10 7 cells/mL). Nude mice (BALB/cSlc- nu/nu , female, 7–8 weeks old; Japan SLC, Shizuoka, Japan) were injected with 1 × 10 6 CD44-negative SK-LMS-1 cells or CD44-positive SK-LMS-1 cells in 5 mg/mL BD Matrigel Matrix (BD Biosciences) in culture medium containing 15% fetal calf serum plus SmGM-2 SingleQuots (Lonza, Basel) at a volume of 100 μL on the left side of the second mammary fat pad. Tumor formation was assessed every day. Tumor volumes were calculated as (L × W × W)/2, where W represents the width, and L represents the length. Statistical analyses were performed on mean tumor volumes at the end of the study using Student’s t -test. ( C ) The upper graph shows the number of mice in which micrometastases were observed in alveoli. The lower graph shows the number of micrometastatic sites found in alveolar sections in each xenografted mouse. It is important to note that in CD44-positive SK-LMS-1 cell-xenografted mice, the frequency of micrometastases in alveolar tissue was significantly higher than that in CD44-negative SK-LMS-1 cell-xenografted mice. Compared with CD44-negative SK-LMS-1 cells, CD44-positive SK-LMS-1 cells exhibit the ability to induce tumor angiogenesis and have higher hematogenous metastatic potential.

Article Snippet: We isolated a candidate population of CD44-positive SK-LMS-1 subclone as human Ut-LMS stem-like cells from human Ut-LMS primary cells and SK-LMS-1 cells, using the CD44-positive selection method with MagCellect Plus Human CD44+ Cell Isolation Kit (R&D Systems, Inc., Minneapolis, MN, USA).

Techniques: Derivative Assay, Injection, Cell Culture

Journal: Current Issues in Molecular Biology

Article Title: Oncological Properties of Intravenous Leiomyomatosis: Involvement of Mesenchymal Tumor Stem-Like Cells

doi: 10.3390/cimb43020084

Figure Lengend Snippet: Differential expression of cyclin B, cyclin E, caveolin 1, Ki-67, LMP2/b1i, and CD44 as potential biomarkers in leiomyoma, leiomyosarcoma, and intravenous leiomyomatosis. ( A ) The photograph shows spindle cell leiomyoma. The low power view (10× field) shows a well-circumscribed tumor nodule in the myometrium composed of broad fascicles of spindle cells. High power view (40× field) shows that uterine leiomyoma (spindle cell) has bland cytological features, with elongated nuclei and fine nuclear chromatin. Immunohistochemistry of leiomyoma tissue was performed using appropriate monoclonal antibodies with standard procedures. ( B ) The photograph shows epithelioid leiomyosarcoma. The low power view (10× field) shows the uterine mass irregular interface with the myometrium, composed of round to polygonal cells with granular eosinophilic cytoplasms. Notice the presence of significant nuclear atypia and clear mitoses. The high power view (40× field) shows that the tumor cells are round to ovoid, with eosinophilic granular cytoplasm and irregularly shaped nuclei. Immunohistochemistry of leiomyoma tissue was performed using appropriate monoclonal antibodies with standard procedures. ( C ) Intravenous leiomyomatosis is a rare neoplasm characterized by nodular masses of histologically benign smooth muscle cells growing within the venous system. Significant smooth muscle hyperplasia was observed in the myometrium. Some tumors invade venous blood vessels. The low power view (10× field) shows no obvious high-grade nuclear atypia or mitotic cell proliferation and necrosis. The high power view (40× field) shows interlaced bundles of homogeneous spindle cells with oval nuclei, eosinophilic cytoplasm, rare mitotic figures, and decorated with several thick-walled small blood vessels, which are consistent with features of leiomyoma. Immunohistochemistry of leiomyoma tissue was performed using appropriate monoclonal antibodies with standard procedures. The areas surrounded by the dotted squares are enlarged.

Article Snippet: We isolated a candidate population of CD44-positive SK-LMS-1 subclone as human Ut-LMS stem-like cells from human Ut-LMS primary cells and SK-LMS-1 cells, using the CD44-positive selection method with MagCellect Plus Human CD44+ Cell Isolation Kit (R&D Systems, Inc., Minneapolis, MN, USA).

Techniques: Expressing, Immunohistochemistry

Journal: Current Issues in Molecular Biology

Article Title: Oncological Properties of Intravenous Leiomyomatosis: Involvement of Mesenchymal Tumor Stem-Like Cells

doi: 10.3390/cimb43020084

Figure Lengend Snippet: CD44-positive uterine mesenchymal tumor stem-like cells in intravenous leiomyomatosis and intravenous uterine leiomyosarcoma. Immunohistochemistry of normal myometrium, uterine leiomyoma, uterine leiomyosarcoma, intravenous leiomyomatosis, and intravenous leiomyosarcoma tissues was performed using appropriate monoclonal antibodies with standard procedures. The five tissue sites were randomly selected: normal myometrium, uterine leiomyoma, uterine leiomyosarcoma, internal tissue of intravenous leiomyomatosis, and intravenous leiomyosarcoma. In a 40× field of view, the positive rates of the six factors were calculated in the five tissue sites under microscopy (Panthera Shimadzu Co. Ltd., Kyoto, Japan); the positive rates at the sites of each tissue are shown in the scatter plot. Int.UL: Intravenous leiomyomatosis, Int.uLMS: Intravenous uterine leiomyosarcoma.

Article Snippet: We isolated a candidate population of CD44-positive SK-LMS-1 subclone as human Ut-LMS stem-like cells from human Ut-LMS primary cells and SK-LMS-1 cells, using the CD44-positive selection method with MagCellect Plus Human CD44+ Cell Isolation Kit (R&D Systems, Inc., Minneapolis, MN, USA).

Techniques: Immunohistochemistry, Microscopy

Journal: Current Issues in Molecular Biology

Article Title: Oncological Properties of Intravenous Leiomyomatosis: Involvement of Mesenchymal Tumor Stem-Like Cells

doi: 10.3390/cimb43020084

Figure Lengend Snippet: The invasion and metastatic process of uterine mesenchymal tumor stem-like cells. The nature of malignant tumors varies from patient to patient. Malignant tumor tissue comprises a heterogeneous cell population, containing many fibroblasts and tumor stem cells in addition to tumor cells. The heterogeneity of malignant tumor cells is one of the main reasons for the difficulty in treating malignant tumors. In particular, malignant tumor stem cells undergo distant metastasis and have resistance to antitumor agents. Understanding the oncological properties of malignant tumor stem cells contributes to the development of new targeted antitumor agents. The complex process of distant metastasis includes detachment, invasion of the tumor microenvironment, and shedding of invasive tumor cells (ITCs) and CD44-positive invasive tumor stem cells (ITSCs) into the bloodstream (intravasation). The majority of ITCs relate to differentiated cancer cells with low tumorigenicity and instead undergo apoptosis. However, CD44-positive ITSCs bear cancer stem cell features, i.e., CD44-positive circulating tumor stem cells (CTSC), survive in the circulating blood, escape from immune surveillance, and go home to secondary sites, extravasate, and ultimately form distant metastatic lesions. The application of cytotoxic drugs for ITSCs and/or CTSCs, such as celecoxib, greatly reduces the incidence of tumor metastases and tumor recurrence . ITC: Invasive tumor cell, ITSC: Invasive tumor stem cell, CTSC: Circulating tumor stem cell.

Article Snippet: We isolated a candidate population of CD44-positive SK-LMS-1 subclone as human Ut-LMS stem-like cells from human Ut-LMS primary cells and SK-LMS-1 cells, using the CD44-positive selection method with MagCellect Plus Human CD44+ Cell Isolation Kit (R&D Systems, Inc., Minneapolis, MN, USA).

Techniques: