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Cultivated cells from mid‐trimester amniotic fluids. IV. Cell type identification via one and two‐dimensional electrophoresis of clonal whole cell homogenates
Author(s) -
Johnston Patricia,
Salk Darrell,
Martin George M.,
Hoehn Holger
Publication year - 1982
Publication title -
prenatal diagnosis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.956
H-Index - 97
eISSN - 1097-0223
pISSN - 0197-3851
DOI - 10.1002/pd.1970020202
Subject(s) - amniotic fluid , isoelectric focusing , biology , gel electrophoresis , polyacrylamide gel electrophoresis , electrophoresis , cell type , isoelectric point , microbiology and biotechnology , cell , cell culture , fetus , genetics , biochemistry , pregnancy , enzyme
Clones of cultivated amniotic fluid cells that have distinct morphologic and growth characteristics (F, AF and E‐type) were examined by one‐dimensional SDS polyacrylamide gel electrophoresis (SDS‐PAGE) and by two‐dimensional electrophoresis employing isoelectric focusing and SDS‐PAGE (IEF‐PAGE). No qualitative differences in band pattern were observed in SDS‐PAGE between the various amniotic fluid cell types, but consistent quantitative differences in the ratios of four bands of presumed filamentous proteins provided good distinction between amniotic fluid cells and postnatal skin fibroblast‐like cells. By adding separation on the basis of electrical charge to that of molecular size (IEF‐PAGE), we observed reproducible qualitative differences in the protein spot patterns between F and both AF and E‐type amniotic fluid cells. At least eight discrete proteins appear not to be synthesized by prenatal F‐type cells in comparison with their isogenic AF and E counterparts under identical culture conditions. The two‐dimensional electrophoretic patterns thus confirm that F and AF amniotic cells, in spite of their morphologic and growth kinetic similarities, are developmentally distinct cell types that retain their differentiated states in culture.