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( Seoung Yob Han ),( Jong Wook Kim ),( Sang Yun Jeon ),( Heung Joong Kim ),( Byung Ock Kim ),( Joo Cheol Park ),( Chang Hoon Song ),( Hyun Seon Jang ) 한국조직공학·재생의학회 2009 조직공학과 재생의학 Vol.6 No.1
Tissue engineering is attracting attention because it is new approaches for the repair and replacement of tissues and organs that have lost function due to variable reasons. Clinical application have already begun. One of the fundamental principles that underlies tissue engineering strategies is that a newly formed tissue must acquire sufficient vascularization. In recent studies, differentiation of the endothelial cells from variable cell sources has been reported, and vascular endothelial growth factor(VEGF) is very important factor in differentiation. This study investigated whether differentiation of BMSC, cultured with VEGF in vitro, into endothelial cell is possible. BMSC was obtained from Dog BMSC was loaded onto an equal volume of 75ul VEGF and incubated at 37oC in 5% CO2 and 95% humidity. Media was changed every 2days. Total RNA was extracted by use of Trizol reagent. The gene expression of cultured cells was investigated by RT-PCR using the primers - nestin, VE-Cadherin, vWF and GAPDH as internal control. And the PCR products were electrophoresed on a 1.2% agarose gel. Digital images of electrophoresd gel were taken and quantified by densitometer(Bio-rad(R)). As time goes by, experiment group show spindle shape more clearly than control group and cell number increase. Nestin marker is expressed in all groups, beside expression of VE-Cadherin and vWF are higher in experiment group than control group. Experiment group treated with VEGF show high expression of the EC-specific marker: VE-Cadherin and vWF. The fact that 5 days culture group reveals the highest level of gene expression means that this group reachs the sufficient differentiation into EC and that it has potential to reduce the time of vascular tissue regeneration.
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