<abstract xmlns="http://www.w3.org/1999/xhtml"> Background: Viral vectors can produce longer-lasting effects than a recombinant protein.Objective: Develop a system of sustained GH1 (a human growth hormone gene) transfer and expression in baby hamster kidney cell (BHK-21) line using recombinant adeno-associated viral vectors pseudotyped with viral capsids from serotype 1 (rAAV2/1).Methods: The expression of GH1 in vitro was examined by reverse transcription polymerase chain reaction and Western blot analysis. Effects of GH1 on cell proliferation were measured by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl-tetrazolium bromide (MTT) experiments. Mice serum IGF-1 and blood glucose, after injection of virus infected BHK-21 cells, were measured by enzyme-linked immunosorbent assays.Results: rAAV2/1-mediated GH1 gene transfer occurred effectively in vitro after 48 hours of transduction at 1 105 vg/mL (multiplicities of infection). MTT experiments indicated notable effects of the GH1 gene on cell proliferation in vitro. Subsequent animal experiment suggested that the injection of virus infected BHK-21 cells might induce increase of serum IGF-1.Conclusion: Our study shows the feasibility of rAAV2/1-mediated GH gene delivery in vitro, applicable for future experimental and clinical investigations. </abstract>

Background: Viral vectors can produce longer-lasting effects than a recombinant protein.Objective: Develop a system of sustained GH1 (a human growth hormone gene) transfer and expression in baby hamster kidney cell (BHK-21) line using recombinant adeno-associated viral vectors pseudotyped with viral capsids from serotype 1 (rAAV2/1).Methods: The expression of GH1 in vitro was examined by reverse transcription polymerase chain reaction and Western blot analysis. Effects of GH1 on cell proliferation were measured by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl-tetrazolium bromide (MTT) experiments. Mice serum IGF-1 and blood glucose, after injection of virus infected BHK-21 cells, were measured by enzyme-linked immunosorbent assays.Results: rAAV2/1-mediated GH1 gene transfer occurred effectively in vitro after 48 hours of transduction at 1 105 vg/mL (multiplicities of infection). MTT experiments indicated notable effects of the GH1 gene on cell proliferation in vitro. Subsequent animal experiment suggested that the injection of virus infected BHK-21 cells might induce increase of serum IGF-1.Conclusion: Our study shows the feasibility of rAAV2/1-mediated GH gene delivery in vitro, applicable for future experimental and clinical investigations.

Human growth hormone gene expression in vitro following rAAV2/1-mediated gene transfer

Department of Clinical Biochemistry, Chinese People’s Liberation Army General Hospital, 28 Fu-Xing Road, Beijing 100853,

Asian Biomedicine

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

Background: Viral vectors can produce longer-lasting effects than a recombinant protein.Objective: Develop a system of sustained GH1 (a human growth hormone...

Human growth hormone gene expression in vitro following rAAV2/1-mediated gene transfer

Article Category: Original article

Online veröffentlicht: 04. Feb. 2017

Seitenbereich: 381 - 387

DOI: https://doi.org/10.5372/1905-7415.0503.056

Schlüsselwörter
Baby hamster kidney cell 21, growth hormone, insulin-like growth factor 1, recombinant adeno-associated virus vector

© 2017

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

Human growth hormone gene expression in vitro following rAAV2/1-mediated gene transfer

Article Category: Original article

Online veröffentlicht: 04. Feb. 2017

Seitenbereich: 381 - 387

DOI: https://doi.org/10.5372/1905-7415.0503.056

SchlüsselwörterBaby hamster kidney cell 21, growth hormone, insulin-like growth factor 1, recombinant adeno-associated virus vector

© 2017

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

Schlüsselwörter
Baby hamster kidney cell 21, growth hormone, insulin-like growth factor 1, recombinant adeno-associated virus vector