Isolation and culture of neuroendocrine cells from fetal rabbit lung using immunomagnetic techniques.
We describe a novel method for the isolation and subsequent culture of pulmonary neuroendocrine cells (PNEC) from normal fetal rabbit lung using immunomagnetic techniques with a monoclonal antibody, MOC-1. This surface antigen has originally been identified on small cell carcinoma of the lung. Our immunohistochemical studies have shown that MOC-1 cross-reacts with PNEC of human and rabbit fetal lungs on frozen sections, and in fixed cultures of rabbit fetal lung. Using a combination of mechanical and enzymatic disaggregation, a single-cell suspension of fetal rabbit lung was obtained. These cells were incubated with MOC-1 conjugated to magnetic beads. PNEC were selectively removed from the heterogeneous mixture using a magnet, giving up to 2-fold enrichment compared with our previously reported method. These cells were maintained in culture in a functional state for up to 7 days. The ability to prepare PNEC from rabbit fetal lung offers an opportunity to develop in vitro models to investigate the physiologic and biochemical properties of these cells, and ultimately it may lead to a better understanding of their function in health and disease.
Speirs
V
V
Department of Pathology, Hospital for Sick Children, Toronto, Ontario, Canada.
Wang
Y V
YV
Yeger
H
H
Cutz
E
E
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D000954:Antigens, Surface / Q000276:immunology; D002460:Cell Line; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D005260:Female; D005333:Fetus; D008168:Lung / Q000737:chemistry / Q000166:cytology* / Q000276:immunology; D008280:Magnetics; D008863:Microspheres; D009490:Neurosecretory Systems / Q000737:chemistry / Q000166:cytology* / Q000276:immunology; D011247:Pregnancy; D011817:Rabbits
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface