We are analyzing https://link.springer.com/article/10.1186/bcr2106.

Title:
Mammosphere culture of metastatic breast cancer cells enriches for tumorigenic breast cancer cells | Breast Cancer Research
Description:
Introduction The identification of potential breast cancer stem cells is of importance as the characteristics of stem cells suggest that they are resistant to conventional forms of therapy. Several techniques have been proposed to isolate or enrich for tumorigenic breast cancer stem cells, including (a) culture of cells in non-adherent non-differentiating conditions to form mammospheres and (b) sorting of the cells by their surface phenotype (expression of CD24 and CD44). Methods We have cultured metastatic cells found in pleural effusions from breast cancer patients in non-adherent conditions without serum to form mammospheres. Dissociated cells from these mammospheres were used to determine the tumorigenicity of these cultures. Expression of CD24 and CD44 on uncultured cells and mammospheres derived from the pleural effusions was documented. Results We found that the majority (20/27) of the pleural effusions tested contained cells capable of forming mammospheres of varying sizes that could be passaged. After dissociation and plating with serum onto adherent dishes, the cells can differentiate, as determined by the increased expression of cytokeratins and MUC1. Analysis of surface expression of CD24 and CD44 on uncultured cells from 21 of the samples showed that the cells from some samples separated into two populations, but some did not. The proportion of cells that could be considered CD44+/CD24low/- was highly variable and did not appear to correlate with the ability to form the larger mammospheres. Of eight pleural effusion mammospheres tested in severe combined immunodeficiency disease (SCID) mice, four were found to induce tumours when only 5,000 or fewer cells were injected, whereas the same number of uncultured cells did not form tumours. The ability to induce tumours appeared to correlate with the ability to produce the larger mammospheres. Uncultured cells from a highly tumorigenic sample (PE14) were uniformly negative for surface expression of both CD24 and CD44. Conclusion This paper shows, for the first time, that mammosphere culture of pleural effusions enriches for cells capable of inducing tumours in SCID mice. The data suggest that mammosphere culture of these metastatic cells could provide a highly appropriate model for studying the sensitivity of the tumorigenic
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cells, mammospheres, cancer, breast, pleural, cell, stem, samples, effusions, article, tumours, expression, mammosphere, culture, pubmed, figure, antibody, tumorigenic, google, scholar, effusion, cas, mice, mammary, uncultured, form, phenotype, injected, stained, analysis, swa, table, size, authors, metastatic, surface, cdcdlow, ability, tumour, produced, data, conditions, cultured, showed, antibodies, population, original, research, number, patients,

Topics {✒️}

cb17/icr-prkdc scid/crl joyce taylor-papadimitriou & joy stem/progenitor cell properties tumour-inducing mammosphere-producing cells epithelial specific antigen-positive article download pdf cancer stem cell full access tumor cells thin grey line thick black line single stem cell human breast cancer isotype-matched labelled controls cancer research uk' cancer research uk phycoerythrin-conjugated ml5 antibody high success rate breast cancer patients full size image breast cancer biology signal-transducing molecule expressed recurrent breast cancer breast cancer campaign fluorescein isothiocyanate breast cancer cells breast cancer res natl cancer inst progenitor cell pool cd44+/cd24low/- phenotype formed mouse mammary gland home office guidelines putative tumorigenic cell differentiated cell lineages tumorigenic 'stem' cells luminal epithelial cells metastatic pleural effusions cell culture technique joyce taylor-papadimitriou normal adult tissues privacy choices/manage cookies pluripotent stem cells neuronal stem cells putative stem cells article number r52 estrogen receptor expression receive oestrogen implants symmetrical cell division cancer metastasis rev nat rev cancer

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         description:The identification of potential breast cancer stem cells is of importance as the characteristics of stem cells suggest that they are resistant to conventional forms of therapy. Several techniques have been proposed to isolate or enrich for tumorigenic breast cancer stem cells, including (a) culture of cells in non-adherent non-differentiating conditions to form mammospheres and (b) sorting of the cells by their surface phenotype (expression of CD24 and CD44). We have cultured metastatic cells found in pleural effusions from breast cancer patients in non-adherent conditions without serum to form mammospheres. Dissociated cells from these mammospheres were used to determine the tumorigenicity of these cultures. Expression of CD24 and CD44 on uncultured cells and mammospheres derived from the pleural effusions was documented. We found that the majority (20/27) of the pleural effusions tested contained cells capable of forming mammospheres of varying sizes that could be passaged. After dissociation and plating with serum onto adherent dishes, the cells can differentiate, as determined by the increased expression of cytokeratins and MUC1. Analysis of surface expression of CD24 and CD44 on uncultured cells from 21 of the samples showed that the cells from some samples separated into two populations, but some did not. The proportion of cells that could be considered CD44+/CD24low/- was highly variable and did not appear to correlate with the ability to form the larger mammospheres. Of eight pleural effusion mammospheres tested in severe combined immunodeficiency disease (SCID) mice, four were found to induce tumours when only 5,000 or fewer cells were injected, whereas the same number of uncultured cells did not form tumours. The ability to induce tumours appeared to correlate with the ability to produce the larger mammospheres. Uncultured cells from a highly tumorigenic sample (PE14) were uniformly negative for surface expression of both CD24 and CD44. This paper shows, for the first time, that mammosphere culture of pleural effusions enriches for cells capable of inducing tumours in SCID mice. The data suggest that mammosphere culture of these metastatic cells could provide a highly appropriate model for studying the sensitivity of the tumorigenic 'stem' cells to therapeutic agents and for further characterisation of the tumour-inducing subpopulation of breast cancer cells.
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            Breast Cancer Stem Cell
            Tumorigenic Cell
            Metastatic Breast Cancer Cell
            Severe Combine Immunodeficiency Disease
            Cancer Research
            Oncology
            Surgical Oncology
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         Breast Cancer Stem Cell
         Tumorigenic Cell
         Metastatic Breast Cancer Cell
         Severe Combine Immunodeficiency Disease
         Cancer Research
         Oncology
         Surgical Oncology
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      name:Breast Cancer Biology Group, King's College London School of Medicine, Guy's Hospital Campus, London, UK
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