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International Orthopaedics

Erschienen in:

01.09.2014 | Original Paper

Nonviral gene transfer into human meniscal cells. Part II: effect of three-dimensional environment and overexpression of human fibroblast growth factor 2

verfasst von: Hsiao-Ping Lee, Ana Rey-Rico, Magali Cucchiarini, Henning Madry

Erschienen in: International Orthopaedics | Ausgabe 9/2014

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Abstract

Purpose

Our aim was to study the effect of three-dimensional (3D) environment and overexpression of human fibroblast growth factor 2 (FGF-2) on meniscal fibrochondrocytes in vitro.

Methods

Human meniscal fibrochondrocytes were transfected with expression plasmid vectors carrying the Photinus pyralis luciferase gene, the Escherichia coli β-galactosidase gene or a human FGF-2 cDNA. Modified fibrochondrocytes were cultivated in 3D alginate hydrogel or cell pellets or in 2D monolayer culture.

Results

The levels of luciferase activity showed a peak at day two and returned to baseline levels by day 11, regardless of the type of cultivation. Both 3D environments supported the secretion of human FGF-2 protein upon FGF-2 transfection. Overexpression of human FGF-2 by genetically modified human meniscal fibrochondrocytes stimulated proliferation but not glycosaminoglycan synthesis only in 3D culture. Culture in alginate spheres resulted in a larger difference in cell numbers compared with pellet cultures.

Conclusions

Three-dimensional alginate spheres are well suited for the culture of genetically modified human meniscal fibrochondrocytes. These data are of value for cell-based approaches to meniscal repair using genetically modified human meniscal fibrochondrocytes overexpressing human FGF-2.

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Titel: Nonviral gene transfer into human meniscal cells. Part II: effect of three-dimensional environment and overexpression of human fibroblast growth factor 2
verfasst von: Hsiao-Ping Lee
Ana Rey-Rico
Magali Cucchiarini
Henning Madry
Publikationsdatum: 01.09.2014
Verlag: Springer Berlin Heidelberg
Erschienen in: International Orthopaedics / Ausgabe 9/2014
Print ISSN: 0341-2695
Elektronische ISSN: 1432-5195
DOI: https://doi.org/10.1007/s00264-014-2405-z

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Abstract

Purpose

Methods

Results

Conclusions

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