Fast separation of macrophages by retention on cross-linked amylose and release by enzymatic amylolysis of the chromatographic material.
Macrophages from mice peritoneal exudate were isolated on basis of specific adherence on epichlorohydrin cross-linked amylose (CLA), a chromatographic gel presenting a high susceptibility to advanced amylolysis with alpha-amylase. The cell suspension, containing predominantly macrophages and lymphocytes, was applied onto the column and incubated for 30 min at 37 degrees C for the adherence of macrophages. After this interval the non-adherent cells were eluted with buffered medium and the CLA support was incubated in the column with an alpha-amylase-buffered solution liquefying the matrix and releasing, in situ, the adherent cell population containing 90% macrophages with a viability higher than 90%.
Desmangles
R J
RJ
Départment de Chimie Université du Québec à Montréal, Canada.
Flipo
D
D
Fournier
M
M
Mateescu
M A
MA
D000688:Amylose / Q000737:chemistry*; D000818:Animals; D002448:Cell Adhesion; D002469:Cell Separation / Q000379:methods*; D002850:Chromatography, Gel / Q000379:methods*; D003432:Cross-Linking Reagents; D004247:DNA / Q000032:analysis; D004996:Ethidium; D005260:Female; D016650:Fluorescein-5-isothiocyanate; D005455:Fluorescent Antibody Technique; D008264:Macrophages / Q000737:chemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D011134:Polysaccharides / Q000032:analysis
D003432:Cross-Linking Reagents; D011134:Polysaccharides; D000688:Amylose; D004247:DNA; D004996:Ethidium; D016650:Fluorescein-5-isothiocyanate