Methodological considerations on Feulgen-staining applied to cells in primoculture: the model of osteoarthritic synovial cells.
Human synovial cells in primoculture are an interesting model for the study of articular joint diseases and anti-rheumatic drugs. Based on results obtained by image cytometry of Feulgen-stained nuclei, we describe the heterogeneity of synovial cell populations and their progression during culture time in primoculture. Using the hydrolysis properties of the Feulgen reaction and their variations dependent on fixatives, we demonstrate the high acid-lability of the condensed chromatin observed in short term cultured nuclei compared to the acid-resistance of decondensed chromatin in long term cultured nuclei; these variations being probably induced by modifications in the molecular supra-organisation of chromatin during the aging of a culture. Finally, due to the cellular heterogeneity of the biological model and its evolution during culture progression, technical compromises are proposed to obtain optimal Feulgen staining, using Böhm-Sprenger fixative and a 1 h hydrolysis by 6 M HCl at 20 degrees C.
Giroud
F
F
Institut Albert Bonniot, Université Joseph Fourier, Faculté de Médecine, La Tronche, France.
Delage
B
B
Monet
J D
JD
Chancel
G
G
Cals
M J
MJ
Ekindjian
O G
OG
D000368:Aged; D000369:Aged, 80 and over; D002478:Cells, Cultured; D002843:Chromatin / Q000235:genetics / Q000378:metabolism; D004396:Coloring Agents; D004261:DNA Replication; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D008297:Male; D008875:Middle Aged; D010003:Osteoarthritis / Q000473:pathology*; D011003:Ploidies; D012394:Rosaniline Dyes; D013194:Staining and Labeling / Q000379:methods*; D013583:Synovial Membrane / Q000473:pathology*; D016707:Tissue Fixation / Q000379:methods*
D002843:Chromatin; D004396:Coloring Agents; C007722:Feulgen stain; D012394:Rosaniline Dyes