Transfection of rabbit articular chondrocytes by the calcium phosphate procedure: Improvement of efficiency and reproducibility.

Cytotechnology
S ViengchareunM Adolphe

Abstract

The understanding of cartilage disorders relies on the possibility of studying mechanisms which monitor the regulation of matrix protein genes through introducing efficiently and in a reproducible manner these genes, or their regulatory regions, into cells. To this end, we attempted to improve the transfection efficiency of rabbit articular chondrocytes by the calcium phosphate procedure. Transfection efficiencies were assessed by measuring the expression of the Lac Z reporter gene encoding β-galactosidase using anin situ staining (X-gal staining) and an enzymatic assay (β-galactosidase assay).Results revealed that addition of 4 U ml(-1) of hyaluronidase before and during transfection increases by 2 to 4-fold the transfection efficiency of rabbit articular chondrocytes. Furthermore, we demonstrated that the use of a "giant" calcium phosphate DNA coprecipitate gives a higher transfection efficiency and much more reproducible results than those obtained with classical small volumes of precipitates.

References

Nov 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·P L FelgnerM Danielsen
Dec 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·W HortonY Yamada
Mar 1, 1971·Clinical Orthopaedics and Related Research·W T Green
Apr 1, 1973·Virology·F L Graham, A J van der Eb

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Citations

May 4, 2004·Methods : a Companion to Methods in Enzymology·Martin Jordan, Florian Wurm
Jan 1, 1997·In Vitro Cellular & Developmental Biology. Animal·S ViengchareunM Adolphe
Mar 21, 2017·Analytical Biochemistry·Mohammad Shahidul MakkiTariq M Haqqi

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