EMAIL THIS PAGE TO A FRIEND

Analytical biochemistry

A fluorometric deoxyribonucleic acid assay for tridimensional lattice cultures of fibroblasts.


PMID 8512073

Abstract

A fast and sensitive in situ assay of deoxyribonucleic acid in miniaturized lattice cultures of fibroblasts is described. Tridimensional collagen and fibrin lattices prepared in 24-well plates were seeded with 50,000 to 200,000 cells. Cultures were fixed with formaldehyde, rinced with isopropanol, and dried. DNA assay was performed directly in the wells by addition of 3,5-diaminobenzoic acid (DABA) reagent. A calibration curve was prepared with calf thymus DNA. Fluorescence of DNA-DABA was evaluated after 45 min incubation (excitation wavelength 420 nm, emission wavelength 490 nm). The method showed linear results from 0.5 to 10 micrograms DNA and proved sensitive for low cell numbers (50,000 per dish). DNA assay in monolayers and in different types of lattices showed that comparable results were obtained in the different models without interference of the extracellular matrix. This technique is regarded as a costless and efficient tool for evaluating the number of cells in lattices in basal conditions or under pharmacological stimulation.

Related Materials

Product #

Image

Description

Molecular Formula

Add to Cart

D12805
3,5-Diaminobenzoic acid, 98%
C7H8N2O2
113832
3,5-Diaminobenzoic acid dihydrochloride, 99%
C7H8N2O2 · 2HCl