Endogenous and exogenous alpha-fetoproteins as differential markers of cultured neonatal mouse Schwann cells and fibroblasts.
alpha-Fetoprotein (AFP) and AFP-gene transcripts were demonstrated in vitro in Schwann (S) cell and fibroblast (F) cell cultures of neonatal mouse origin. All S and F cells of primary cultures and of established cell lines expressed the AFP gene. AFP mRNA was detected by an in situ hybridization technique using a 35S-AFP-cDNA probe. AFP was localized by immunocytoperoxidase labelling using purified anti-AFP antibodies. The amounts of stained endogenous AFP, estimated semiquantitatively, were about 3-fold higher in S cells than in F cells. After incubating the cultures with exogenous mouse AFP, both S and F cells showed significant ability to take up the protein; the amount of internalized protein was found to be higher in F cells than in S cells. Moreover, the uptake of AFP fluorescein conjugates (FITC-AFP), estimated quantitatively by fluorometry, also gave higher values for F cells. The cytoplasm of F cells exhibited a characteristic fluorescence pattern, strongly illuminated and dispersed grains; the cytoplasm of S cells was regularly labelled. If exogenous FITC-AFP uptake could be used to distinguish labelled F cells from S cells (with application for identification and selection of F cells), the immunocytochemically stained endogenous AFP could allow S cells to be distinguished from F cells (using the dilutions of antibodies still staining the S cells but which lead to the absence of F-cell labelling). The two procedures, which can be used independently or together, may constitute differential markers for S cell and F cultures in, i.e., nerve regeneration of neurofibroma studies using the model of mixed S and F culture also containing other types of cells.
Trojan
J
J
Laboratoire de Chimie des Proteines, CNRS, Villejuif, France.
Boutry
J M
JM
Hauw
J J
JJ
Lafarge-Frayssinet
C
C
Deugnier
M A
MA
Hajeri-Germond
M
M
Abramsky
O
O
Uriel
J
J
D000818:Animals; D000831:Animals, Newborn / Q000378:metabolism*; D015415:Biomarkers; D002478:Cells, Cultured; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D016650:Fluorescein-5-isothiocyanate; D005455:Fluorescent Antibody Technique; D015870:Gene Expression / Q000187:drug effects; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D051379:Mice; D008815:Mice, Inbred Strains; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D012583:Schwann Cells / Q000378:metabolism*; D000509:alpha-Fetoproteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology
D015415:Biomarkers; D012333:RNA, Messenger; D000509:alpha-Fetoproteins; D016650:Fluorescein-5-isothiocyanate