Cellulase slide preparation of methylcellulose cultures of hemopoietic cells.
A technique is described for making slides of colonies grown in methylcellulose semisolid medium. The entire contents of a 0.5-ml culture in a 24-well plate is digested using 0.25 ml of a 1.08-mg/ml solution of cellulase dissolved in fetal calf serum. The methylcellulose is digested overnight, and the well contents are transferred to slides and are stained. Adherent colonies are recovered using a fetal calf serum solution containing trypsin and EDTA to flush any residue from the well; the recovered cells are transferred to slides and then stained. This method produces slides free of the darkly stained methylcellulose background that interferes with viewing colonies and studying individual cells.
D000818:Animals; D001854:Bone Marrow Cells; D002478:Cells, Cultured; D002480:Cellulase; D004285:Dogs; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D008264:Macrophages / Q000166:cytology; D008747:Methylcellulose
D008747:Methylcellulose; D002480:Cellulase