The experiment study of mesenchymal stem cells modified wtih galectin- 1 gene co-cultured with dermal matrix patch

Abstract

Objective To explore the feasibility of mesenchymal stem cells(MSCs) modified wtih galectin- 1 gene co- cultured with dermal matrix patch. Methods After the bone marrow MSCs(BMSCs) were transfected with Ad- green fluorescent protein(GFP)- galectin- 1, the fluorescence in BMSCs was observed under fluorescence microscope.Real- time quantitative reverse transcriptase- polymerase chain reaction(RT- qPCR) was used to detect the expression of galectin- 1 gene, and the protein expression of galectin- 1 and desmin was detected by enzyme linked immune sorbent assay(ELISA)and Western blotting, respectively.The growth of BMSCs transfected with Ad- GFP- galectin- 1 and co- cultured with acellular dermal matrix patch(ADM) was observed. Results The green fluoresence was observed and reached to peak at 72 h after the transfection of BMSCs with Ad- GFP- galectin- 1 under fluorescence microscope.The result of RT- qPCR and ELISA confirmed that BMSCs over- expressed galectin- 1.The RT- qPCR result of galectin- 1,which the relative expression folds was 1.00 in phosphate buffer(PBS) group,4.11 in Ad- GFP- galectin- 1 group,1.09 in Ad- GFP group;The result of galectin- 1 measured by ELISA was that 48 h:(197.28±8.78),(25.52±1.56),(24.53±2.96)μg/L; 72 h:(264.33± 5.43),(25.40±2.06),(25.46±1.13)μg/L;96 h:(172.41±3.61),(25.22±1.61),(28.01±1.26) μg/L; 120 h:(155.53±7.25),(24.84±2.35),(26.65±1.54)μg/L ordered by Ad- GFP-galectin- 1 group, Ad- GFP group and PBS group.The expression of desmin which co- cultured with dermal matrix patch(ADM) were well grown. Conclusion Galectin- 1 could induce BMSCs to express desmin.BMSCs transfected with Ad- GFP- galectin- 1 showed good compatibility with ADM. Key words: Galectin-1; Bone mesenchymal marrow cell; Acellular dermal matrix patch; Co-culture