PMC:17827 / 24416-25301 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/17827","sourcedb":"PMC","sourceid":"17827","source_url":"http://www.ncbi.nlm.nih.gov/pmc/17827","text":"Negative isolation from primary culture\nTrypsinized and washed RA and OA synovial cells from primary culture (107/ml) were incubated with 4 × 107/ml Dynabeads® M-450 CD14 (clone RMO52; Dynal) in PBS/2% FCS for 1 h at 4°C under bidirectional rotation. Nine milliliters of PBS/2% FCS were then added and the conjugated cells collected using the Dynal magnetic particle concentrator®. Magneto-bead-conjugated cells and unconjugated cells were collected and washed twice in PBS/2% FCS; cell composition and phenotype were analyzed by flow cytometry using the antibodies presented in Table 3. For the purpose of comparison with fourth-passage cells obtained by conventional isolation (ie repeated passaging; see later), negatively isolated RA-SFB and OA-SFB were then passaged four times by culture in complete DMEM/10% FCS (see earlier) with a 1:3 split of confluent cells in each passage.","divisions":[{"label":"Title","span":{"begin":0,"end":39}}],"tracks":[]}