Construction of recombinant adenoviral vector to coexpress human neurotrophin3 and EGFP gene and its conduction efficiency to rat cochlea in vitro.
- Author:
Bo DU
1
;
Ping WANG
;
Baodong DU
Author Information
1. Department of Otolaryngology, First Hospital of Jilin University, Changchun 130021, China.
- Publication Type:Journal Article
- MeSH:
Adenoviridae;
genetics;
Animals;
Basilar Membrane;
cytology;
Cell Survival;
genetics;
Cells, Cultured;
Cochlea;
cytology;
Genetic Vectors;
Hair Cells, Auditory;
cytology;
Humans;
Neurotrophin 3;
genetics;
Rats;
Rats, Inbred F344;
Transfection
- From:
Journal of Clinical Otorhinolaryngology Head and Neck Surgery
2008;22(10):462-465
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To construct an adenoviral vector that codes for both human NT3 and EGFP, to confirm the transduction efficiency in rat cochlear cultures and to assess the protection of NT3 on SGNs survival.
METHOD:PAdeasy-1 and pAdTrack CMV were used to constructed Ad/NT3 adenovirus and then to transfer postnatal day 3 rat cochlear cultures. The transduction efficiency was determined by microscope observation. The amounts of SGNs were counted to evaluated protection of Ad/NT3 on SGNs survival.
RESULT:EGFP positive cells were observed in all cochlear turns. There was approximately 49% in outer sulcus cells and 27% in the interdental cells; less than 2% of the hair cells and SGN. The amounts of SGN of treated Ad/NT3 adenovirus are more than cochlea SGN only Ad/EGFP adenovirus after cultured for 15 days.
CONCLUSION:Ad/NT3 adenovirus could transduce EGFP and NT3 in large number of supporting cells, but few hair cells or SGNs. The putative release of NT3 from these supporting cells could enhance cell survival and promote neurite outgrowth from SGNs.
