[Expression of exogenous gene in bone marrow stromal cells by liposome].
OBJECTIVE
To transform eukaryotic expression vector pEGFP-PDX-1 into marrow stromal cells by liposome and to optimize the conditions of transformation.


METHODS
The recombinant vector was identified by enzyme digestion analysis and sequencing. The recombinant plasmid was transformed into bone marrow stromal cells and it changed the quantity of DNA or liposome. The expression of PDX-1 gene in the transformed cells was detected by immunocytochemical staining.


RESULTS
Enzyme digestion analysis and sequencing showed that the interesting gene was integreted into the recombinant vector. We obtained satisfactory efficiency of transfection when the ratio of DNA and liposome was 1 : 1 or 1 : 2. The PDX-1 in the transformed cells was expressed by immunocytochemical staining.


CONCLUSIONS
The eukaryotic expression vector pEGFP-PDX-1 was constructed for the first time in China. We have enhanced the efficiency of transfection by optimizing the transformation conditions. It is possible to use the bone marrow stromal cells as seed cells in tissue-engineering.
Sun
Ji-ping
JP
Department of Pediatrics, Xiangya Hospital, Central South University, Changsha 410008, China.
Jia
Yan-jie
YJ
Luo
Fang
F
Song
Jian-hui
JH
Yang
Yu-jia
YJ
D000818:Animals; D001483:Base Sequence; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism*; D002478:Cells, Cultured; D005057:Eukaryotic Cells / Q000378:metabolism; D018398:Homeodomain Proteins / Q000096:biosynthesis* / Q000235:genetics; D008081:Liposomes; D008297:Male; D008969:Molecular Sequence Data; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism*; D023822:Tissue Engineering; D015534:Trans-Activators / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection
D018398:Homeodomain Proteins; D008081:Liposomes; D011994:Recombinant Proteins; D015534:Trans-Activators; C084010:pancreatic and duodenal homeobox 1 protein