Glycosphingolipid patterns in primary mouse kidney cultures
Primary kidney cultures from C57BL/6J mice, 6 weeks of age or older, were produced using D-valine medium to select for epithelial cell growth. After allowing the cells to attach and proliferate for 1 week following plating, medium was changed once per week. Cells formed nearly confluent monolayers during the second week of culture. The cultured cells contained all of the glycosphingolipids seen in the adult kidney, analyzed by high performance liquid chromatography as their perbenzoyl derivatives. Glucosylceramide, however, was highly predominant in the cultured cells, whereas dihexosyl- and trihexosylceramides predominate in the intact kidney. Sex differences in glycolipid contents found in the intact kidney were also apparent in these cultured cells: The concentration of neutral glycolipids, in general, was higher in male cells than in those derived from females, and the male-specific glycolipid nonhydroxy fatty acid digalactosylceramide was high in male cells but very low in female cells. Neutral glycosphingolipids were labeled in 2-week-old cultures using (/sup 3/H)palmitate. The (/sup 3/H)palmitate was incorporated into all of the glycolipids within 2 hr of labeling. Hence, adult mouse kidney cells in D-valine medium retain their differentiated characteristics for a sufficient period of time to allow investigation of glycolipid syntheses in monolayer cultures of epithelial cells derived from this organ.
- Research Organization:
- E.K. Shriver Center, Waltham, MA
- OSTI ID:
- 6528024
- Journal Information:
- J. Cell. Physiol.; (United States), Vol. 129:3
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
GLYCOLIPIDS
BIOSYNTHESIS
TRITIUM COMPOUNDS
AGE DEPENDENCE
BIOCHEMISTRY
CELL CULTURES
HEXADECANOIC ACID
KIDNEYS
LIQUID COLUMN CHROMATOGRAPHY
MICE
SEX DEPENDENCE
VALINE
AMINO ACIDS
ANIMALS
BODY
CARBOHYDRATES
CARBOXYLIC ACIDS
CHEMISTRY
CHROMATOGRAPHY
LABELLED COMPOUNDS
LIPIDS
MAMMALS
MONOCARBOXYLIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANS
RODENTS
SACCHARIDES
SEPARATION PROCESSES
SYNTHESIS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques