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001-es BibID:	BIBFORM013301
Első szerző:	Juhász Tamás (biológus, orvosbiológus)
Cím:	Optimalized transient transfection of chondrogenic primary cell cultures / Juhász Tamás, Matta Csaba, Mészár Zoltán, Nagy Georgina, Szíjgyártó Zsolt, Molnár Zsanett, Kolozsvári Bernadett, Bakó Éva, Zákány Róza
Dátum:	2010
ISSN:	1895-104X
Megjegyzések:	We aimed to find a transfection method which provides high efficiency with minimal cytotoxic and/or apoptotic effects for gene transfer into multilayer primary chondrogenic cell cultures. The pEGFP-C1 plasmid was introduced into the cell culture and the efficiency of transformation quantified by GFP fluorescence; the resulting nucleofection was effective but resulted in severe apoptosis. Two liposomal reagents designed to allow transfection into adherent cells did not deliver the plasmids sufficiently and cartilage formation did not occur.In addition, a third liposomal compound, recommended for transfection into either adherent or suspension cell cultures, lead to acceptable transfection efficiency but no cartilage formation. When an amphiphilic reagent was used however, there was acceptable transfection efficiency as well as cartilage formation. The viability of the cells which were transfected using the amphiphilic reagent remained unaffected but proliferation was severely diminished, particularly in the presence of GFP. In addition, the amount of cartilage decreased when GFP was expressed, despite unchanged levels of mRNAs of sox9 and aggrecan core protein, factors reflecting on the efficiency of chondrogenesis. Overexpression of both the constitutively active delta and gamma isoforms of catalytic subunit of calcineurin, a protein phosphatase described as a positive regulator of chondrogenesis, decreased protein level of Sox9 and subsequent cartilage formation. In conclusion, we found that amphiphilic reagent applied prior to the adhesion of cells provides a useful means to transfer plasmids to primary differentiating chondrogenic cells.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban Cartilage differentiation Mesenchymal cell culture Chondrifying primary cells Transfection efficiency Calcineurin overexpression
Megjelenés:	Central European Journal of Biology. - 5 : 5 (2010), p. 572-584. -
További szerzők:	Matta Csaba (1980-) (molekuláris biológus, genetikus, angol szakfordító) Mészár Zoltán Mihály (1977-) (agrármérnök) Nagy Georgina (1980-) (orvosbiológus) Szíjgyártó Zsolt (1978-) (vegyész) Molnár Zsanett (1982-) (analitikus) Kolozsvári Bernadett (1982-) (molekuláris biológus, genetikus) Bakó Éva (1958-) (biokémikus) Zákány Róza (1963-) (anatómus-, kötőszövetbiológus)
Internet cím:	Szerző által megadott URL DOI Intézményi repozitóriumban (DEA) tárolt változat
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