Development of pulmonary neuroendocrine cells of fetal hamster in explant culture.
Fetal hamster lung explant was cultured in serum-free medium on gestational Day 11-2 days before the appearance of pulmonary neuroendocrine cells (PNEC)--and the development and differentiation of PNEC from immature fetal lung epithelium was examined through immunostaining for neural cell adhesion molecule (NCAM) to establish an in vitro system to study the mechanisms involved. PNEC were present in the main bronchus after 2 days of culture. Thereafter, NCAM-positive clusters of PNEC increased and were distributed from the large bronchus to the terminal bronchiole with a proximal-to-distal wave. To elucidate the role of NCAM in the fetal development of PNEC, whole fetal lung was cultured on gestational Day 11 with an anti-NCAM antibody. This antibody slightly inhibited the growth and branching morphogenesis of the lung and disturbed the formation of PNEC clusters. NCAM may function to form clusters of PNEC known as neuroepithelial bodies. We cultured fetal lung epithelial explant at gestational Day 11 after removing mesenchyme, including nerve tissue, with dispase digestion. Immunohistochemical staining for NCAM revealed that PNEC were induced in cultured fetal epithelium without mesenchymal tissue, but basement membrane Matrigel was necessary to maintain cultured epithelium. In conclusion, PNEC derive from immature airway epithelial cells. This organ culture system, therefore, is a useful experimental model and should facilitate further investigations of the development and differentiation of PNEC. Mesenchymal and neural tissues are not always necessary for the development of PNEC, but matrix substance and/or growth factors may be required to induce or maintain PNEC.
Ito
T
T
Department of Pathology, Yokohama City University School of Medicine, Japan.
Nogawa
H
H
Udaka
N
N
Kitamura
H
H
Kanisawa
M
M
D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D002454:Cell Differentiation / Q000276:immunology; D002469:Cell Separation; D006224:Cricetinae; D004847:Epithelial Cells / Q000166:cytology / Q000276:immunology; D005260:Female; D008168:Lung / Q000166:cytology* / Q000196:embryology* / Q000276:immunology; D008297:Male; D019006:Neural Cell Adhesion Molecules / Q000276:immunology; D009490:Neurosecretory Systems / Q000166:cytology* / Q000196:embryology* / Q000276:immunology; D009924:Organ Culture Techniques
D000906:Antibodies; D019006:Neural Cell Adhesion Molecules