Cultures of human Schwann cells isolated from fetal nerves.
We describe a technique for the preparation of highly purified populations of Schwann cells (SC) from human fetal nerves. Cultures were prepared by chemical and mechanical dissociation of human fetal sciatic nerves by modification of the method of Kreider et al. developed for newborn rat nerve. A time course analysis of some SC-associated markers at different times in vitro was performed employing immunofluorescence (IF) and immunoperoxidase (IP) to determine the percentage of SC in culture and to evaluate the maintenance of specific SC characteristics. We compared this method with that of Askanas et al. which produces enriched SC cultures by utilizing successive re-explantation of the original nerve explant. After 48 h, approximately 90% of the cells were bipolar and S-100+ and over the next two weeks about 70-80% of cells were SC by cytologic and immunocytologic criteria. At 35 days, 35% were SC, whereas less than or equal to 2.5% of 35-day-old multi-explant cultures were SC. The SC obtained by this method displayed the typical morphological and immunological characteristics: they expressed surface laminin and nerve growth factor receptors, whereas fibronectin, which is localized on fibroblast surface, was absent.
Scarpini
E
E
Department of Neurology, Dino Ferrari Center for Neuromuscular Diseases, University of Milan, Italy.
Kreider
B Q
BQ
Lisak
R P
RP
Meola
G
G
Velicogna
M E
ME
Baron
P
P
Beretta
S
S
Buscaglia
M
M
Ross
A H
AH
Scarlato
G
G
D002469:Cell Separation; D002478:Cells, Cultured; D005267:Femoral Nerve / Q000166:cytology* / Q000196:embryology; D005333:Fetus; D005353:Fibronectins / Q000032:analysis; D006801:Humans; D007797:Laminin / Q000032:analysis; D011956:Receptors, Cell Surface / Q000032:analysis; D017475:Receptors, Nerve Growth Factor; D009418:S100 Proteins / Q000032:analysis; D012583:Schwann Cells / Q000166:cytology* / Q000378:metabolism; D013997:Time Factors
D005353:Fibronectins; D007797:Laminin; D011956:Receptors, Cell Surface; D017475:Receptors, Nerve Growth Factor; D009418:S100 Proteins