Home LiteratureArticle Details
PMID: 873561 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Fibroblast monolayer cultures in scintillation counting vials: metabolic and growth experiments using radioisotopes and a microfluoremetric DNA assay.

In vitro ·Vol. 13 ·No. 4 ·1977-04-00 ·Pages 237-44

Fujimoto WY, Teague J, Williams RH

Abstract

We have developed a simple technique for the investigation of cellular metabolism and growth in cultured human fibroblasts which facilitates experiments using up to 3 x 10(5) cells in each of 100 or more culture vessels. The method has been used to study cell growth, glucose utilization and oxidation, and protein, RNA and DNA sysnthesis. The use of radiolabeled substrates in tracer experiments is simplified since transfer of cell material is not required. Methods for measuring both total cellular protein and DNA have been adapted to this culture system. Although we have used this tecnique for fibroblast cultures, it also can be easily applied to experiments on any other type of cell that can be grown in a monolayer.

MeSH Terms
Carbon Radioisotopes Cell Division Cells, Cultured Cycloheximide/pharmacology DNA/biosynthesis Dactinomycin/pharmacology Fibroblasts/metabolism Fluorometry/methods Glucose/metabolism Humans Mitomycins/pharmacology Protein Biosynthesis RNA/biosynthesis Skin/embryology Tritium
Chemicals
Carbon Radioisotopes Mitomycins Tritium Dactinomycin RNA DNA Cycloheximide Glucose
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Fujimoto W Y
Teague J
Williams R H
References (15)
15 references, click to expand
  1. The division cycle and RNA-synthesis in diploid human cells at different passage levels in vitro.
    Exp Cell Res. 1966 Jun;42(3):673-84 PMID: 5922078
  2. The fluorometric measurement of deoxyribonucleic acid in animal tissues with special reference to the central nervous system.
    J Biol Chem. 1958 Jul;233(1):184-8 PMID: 13563465
  3. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid.
    Biochem J. 1956 Feb;62(2):315-23 PMID: 13293190
  4. An improved fluorometric assay for DNA.
    Anal Biochem. 1971 Jan;39(1):197-201 PMID: 5544593
  5. A method for the determination of desoxyribonucleic acid, ribonucleic acid, and phosphoproteins in animal tissues.
    J Biol Chem. 1945;161:83-9 PMID: 21005717
  6. Cell size and nucleic acid content in the diploid human cell line WI-38 during aging.
    Med Exp Int J Exp Med. 1969;19(6):313-20 PMID: 5408801
  7. Insulin action on the cultured human fibroblast. Glucose uptake, protein synthesis, RNA synthesis.
    Diabetes. 1974 May;23(5):443-8 PMID: 4830179
  8. A novel method for measuring incorporation of radioactive precursors into nucleic acids and proteins of cells in monolayer culture.
    Anal Biochem. 1972 Mar;46(1):101-7 PMID: 5017648
  9. Alteration in cellular RNAs during the in vitro lifespan of cultured human diploid fibroblasts.
    Cell. 1975 Oct;6(2):179-84 PMID: 1182799
  10. Rapid filter method for the microfluorometric analysis of DNA.
    Anal Biochem. 1975 Dec;69(2):572-82 PMID: 1240734
  11. Leakage of cyclic AMP from human diploid fibroblasts in tissue culture.
    Nat New Biol. 1973 Nov 28;246(152):119-20 PMID: 4357046
  12. Protein measurement with the Folin phenol reagent.
    J Biol Chem. 1951 Nov;193(1):265-75 PMID: 14907713
  13. A modified fluorometric method for the determination of microgram quantities of DNA from cell or tissue cultures.
    Anal Biochem. 1976 Mar;71(1):313-7 PMID: 179442
  14. Abnormalities in proliferation and protein synthesis in skin fibroblast cultures from patients with diabetes mellitus.
    Diabetes. 1977 Apr;26(4):284-90 PMID: 849809
  15. GROWTH AND GLYCOLYSIS IN THE HUMAN DIPLOID CELL STRAIN WI-38.
    Proc Soc Exp Biol Med. 1965 Apr;118:1109-13 PMID: 14277674
Article Info
Journal
In vitro
Abbr.
In Vitro
ISSN
0073-5655
Published
1977-04-00
Pages
237-44
Language
English
Region
United States
NLM ID
0063733
Subset
IM
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]