Transferring gene to rabbit corneal endothelial cells in vitro by cationic polymer
- VernacularTitle:阳离子聚合物介导的体外培养兔角膜内皮细胞基因转移
- Author:
Lin ZHANG
;
Zhaoqin HAO
- Publication Type:Journal Article
- Keywords:
corneal endothelial cells;
cationic polymer;
gene transfer;
Na+-K+-ATPase
- From:
Journal of Xi'an Jiaotong University(Medical Sciences)
1981;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To observe the transfection efficiency of transferring plasmids pEGFP-N1 and pIRE-EGFP encoding enhanced green fluorescent protein(EGFP) to rabbit corneal endothelial cells(RCECs) by cationic polymer and the change of cell activity by detecting the activity of Na+-K+-ATPase.Methods RCEC was cultured by digestion and identification by neurone specific enolase(NSE) stain.Sofast TM mediated plasmid pEGFP-N1 and pIRE-EGFP to transfect RCECs and we compared the transfection efficiency at different time and in different groups by calculating the number of fluorescent cells.We observed the activity change of transfection and non-transfection RCEC by detecting the activity of Na+-K+-ATPase.Results After pEGFP-N1 and pIRE-EGFP transfected RCEC with Sofast TM by different ratio,RCEC expressed EGFP in 24-48 h.After transfection 48 h the group of SofastTM∶pEGFP-N1=3.2∶1 had the max transfection efficiency 3.6% and the max transfection efficiency of group SofastTM∶pIRE-EGFP =3.2∶1 was 3.5%,at 24 h after transfection.There were no obvious differences between the transfection and non-transfection groups of cell number and activity of Na+-K+-ATPase.Conclusion Sofast TM can mediate exogenous gene trasfering to RCEC effectively.pEGFP-N1 and pIRE-EGFP are the suitable non-virus vectors for gene transferring to RCECs.
