Pure populations of murine macrophages from cultured embryonic stem cells. Application to studies of chemotaxis and apoptotic cell clearance.
Embryonic stem cells provide a potentially convenient source of macrophages in the laboratory. Given the propensity of macrophages for plasticity in phenotype and function, standardised culture and differentiation protocols are required to ensure consistency in population output and activity in functional assays. Here we detail the development of an optimised culture protocol for the production of murine embryonic stem cell-derived macrophages (ESDM). This protocol provides improved yields of ESDM and we demonstrate that the cells are suitable for application to the study of macrophage responses to apoptotic cells. ESDM so produced were of higher purity than commonly used primary macrophage preparations and were functional in chemotaxis assays and in phagocytosis of apoptotic cells. Maturation of ESDM was found to be associated with reduced capacity for directed migration and increased capacity for phagocytic clearance of apoptotic cells. These results show ESDM to be functionally active in sequential phases of interaction with apoptotic cells and establish these macrophage populations as useful models for further study of molecular mechanisms underlying the recognition and removal of apoptotic cells.
University of Edinburgh/MRC Centre for Inflammation Research, Queen's Medical Research Institute, 47 Little France Crescent, Edinburgh, EH16 2TJ, UK.
Lihui
L
Zhuang
John D
JD
Pound
Jorine J L P
JJ
Willems
A Helen
AH
Taylor
Lesley M
LM
Forrester
Christopher D
CD
Gregory
D000818:Animals; D015703:Antigens, CD / Q000276:immunology; D000945:Antigens, Differentiation, T-Lymphocyte / Q000276:immunology; D017209:Apoptosis / Q000276:immunology; D039481:CD11b Antigen / Q000276:immunology; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002460:Cell Line; D002478:Cells, Cultured; D002633:Chemotaxis / Q000276:immunology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000276:immunology; D005434:Flow Cytometry; D016130:Immunophenotyping; D008264:Macrophages / Q000166:cytology* / Q000276:immunology; D051379:Mice; D010587:Phagocytosis / Q000276:immunology; D015203:Reproducibility of Results; D013997:Time Factors
D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D039481:CD11b Antigen; C062279:CD6 antigen