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of fetal liver-derived macrophages (FLDMs)\nFetal livers were excised from embryos at E12.5 and E13.5, respectively, washed in PBS and dissociated enzymatically for 60 min at 37°C. The digestion buffer (150 μl per liver) comprised 0.6 U/ml dispase I (Roche), 0.1% collagenase D (Roche), 10 U DNase (Roche), and 20% FCS in PBS. X-Vivo 15 medium (Cambrex, East Rutherford, USA) was added to the resulting cell suspension, and after centrifugation (200 × g; 3 min) cells were resuspended in X-Vivo 15 medium supplemented with 50 ng/ml macrophage colony-stimulating factor (M-CSF; Sigma-Aldrich, St. Louis, USA) and cultured on non-treated tissue-culture dishes at 37°C with 5% CO2. Every second or third day the medium was changed by centrifugation. Following withdrawal of M-CSF on day 6 after excision, adherent cells were cultured for an additional 24-48 h in X-Vivo 15 medium."}

    craft-ca-core-ex-dev

    {"project":"craft-ca-core-ex-dev","denotations":[{"id":"T16445","span":{"begin":21,"end":26},"obj":"UBERON:0002107"},{"id":"T16446","span":{"begin":35,"end":46},"obj":"CL:0000235"},{"id":"T16447","span":{"begin":61,"end":67},"obj":"UBERON:0002107"},{"id":"T16448","span":{"begin":86,"end":93},"obj":"UBERON:0000922"},{"id":"T16449","span":{"begin":158,"end":171},"obj":"CHEBI_GO_SO_EXT:enzyme"},{"id":"T16450","span":{"begin":206,"end":212},"obj":"CHEBI_CHMO_EXT:buffer_solution"},{"id":"T16451","span":{"begin":225,"end":230},"obj":"UBERON:0002107"},{"id":"T16452","span":{"begin":303,"end":308},"obj":"GO_EXT:0004536"},{"id":"T16453","span":{"begin":414,"end":418},"obj":"CL_GO_EXT:cell"},{"id":"T16454","span":{"begin":473,"end":478},"obj":"CL_GO_EXT:cell"},{"id":"T16455","span":{"begin":543,"end":553},"obj":"CL:0000235"},{"id":"T16456","span":{"begin":543,"end":579},"obj":"PR_EXT:000005930"},{"id":"T16457","span":{"begin":581,"end":582},"obj":"CL:0000235"},{"id":"T16458","span":{"begin":581,"end":586},"obj":"PR_EXT:000005930"},{"id":"T16459","span":{"begin":647,"end":653},"obj":"UBERON:0000479"},{"id":"T16460","span":{"begin":685,"end":688},"obj":"CHEBI:16526"},{"id":"T16461","span":{"begin":782,"end":787},"obj":"PR_EXT:000005930"},{"id":"T16462","span":{"begin":813,"end":821},"obj":"GO_PATO_EXT:biological_adhesion_or_attachedness"},{"id":"T16463","span":{"begin":822,"end":827},"obj":"CL_GO_EXT:cell"}],"text":"Preparation of fetal liver-derived macrophages (FLDMs)\nFetal livers were excised from embryos at E12.5 and E13.5, respectively, washed in PBS and dissociated enzymatically for 60 min at 37°C. The digestion buffer (150 μl per liver) comprised 0.6 U/ml dispase I (Roche), 0.1% collagenase D (Roche), 10 U DNase (Roche), and 20% FCS in PBS. X-Vivo 15 medium (Cambrex, East Rutherford, USA) was added to the resulting cell suspension, and after centrifugation (200 × g; 3 min) cells were resuspended in X-Vivo 15 medium supplemented with 50 ng/ml macrophage colony-stimulating factor (M-CSF; Sigma-Aldrich, St. Louis, USA) and cultured on non-treated tissue-culture dishes at 37°C with 5% CO2. Every second or third day the medium was changed by centrifugation. Following withdrawal of M-CSF on day 6 after excision, adherent cells were cultured for an additional 24-48 h in X-Vivo 15 medium."}

    craft-ca-core-dev

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