Culture of fetal alveolar epithelial type II cells in serum-free medium
| Autor: | Caroline Fraslon, GaËlle Rolland, Jacques R. Bourbon, Michel Rieutort, Cécile Valenza |
|---|---|
| Rok vydání: | 1991 |
| Předmět: |
Clinical Biochemistry
Biotin Biology Culture Media Serum-Free Epithelium Choline chemistry.chemical_compound Selenium Sodium pyruvate Fetus Pulmonary surfactant Biosynthesis Pregnancy Phosphatidylcholine Pyruvic Acid Animals Microscopy Phase-Contrast Pyruvates Cells Cultured Phospholipids chemistry.chemical_classification Transferrin Epithelial Cells Rats Inbred Strains Cell Biology General Medicine DNA Rats Pulmonary Alveoli Chemically defined medium Microscopy Electron chemistry Biochemistry Microscopy Fluorescence Cell culture Phosphatidylcholines Female Fetal bovine serum Cell Division Developmental Biology Thymidine |
| Zdroj: | In vitro cellulardevelopmental biology : journal of the Tissue Culture Association. (11) |
| ISSN: | 0883-8364 |
| Popis: | A serum-free culture medium (defined medium = DM) was elaborated by adding to Eagle's minimum essential medium (MEM), non-essential amino acids, transferrin, putrescine, tripeptide glycyl-histidyl-lysine, somatostatin, sodium selenite, ethanolamine, phosphoethanolamine, sodium pyruvate, and metal trace elements. This medium was tested for its ability to support sustained surfactant biosynthesis in fetal alveolar epithelial type II cells. For up to 8 days, ultrastructure was maintained with persistence of lamellar inclusion bodies. Thymidine incorporation into DNA was enhanced about 50% in DM as compared with MEM, whereas it was enhanced 300% in 10% fetal bovine serum. With DM, the incorporation of tritiated choline into phosphatidylcholine (PC) of isolated surfactant material was about twice that with MEM. Deletion experiments evidenced the prominent role of pyruvate, transferrin, and selenium in the stimulation of surfactant PC biosynthesis. The addition of biotin to DM enhanced surfactant PC biosynthesis slightly and nonsurfactant PC biosynthesis markedly. The presence of nucleosides seemed unfavorable to the synthesis of surfactant PC. Type II cells responded to the addition of epidermal growth factor and insulinlike growth factor-I both by increased thymidine incorporation into DNA and choline incorporation into PC. It is concluded that DM represents a useful tool for cultivating type II cells without loss of their specialized properties and for studying the regulation of cell proliferation and surfactant biosynthesis in a controlled environment. |
| Databáze: | OpenAIRE |
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