Genetic modification of embryonic stem cells with VEGF enhances cell survival and improves cardiac function

Xiaoyan Xie, Feng Cao, Ahmad Y. Sheikh, Zongjin Li, Andrew J. Connolly, Xuetao Pei, Ren Ke Li, Robert C. Robbins, Joseph C. Wu

Research output: Contribution to journalArticlepeer-review

43 Scopus citations

Abstract

Cardiac stem cell therapy remains hampered by acute donor cell death posttransplantation and the lack of reliable methods for tracking cell survival in vivo. We hypothesize that cells transfected with inducible vascular endothelial growth factor 165 (VEGF165) can improve their survival as monitored by novel molecular imaging techniques. Mouse embryonic stem (ES) cells were transfected with an inducible, bidirectional tetracycline (Bi-Tet) promoter driving VEGF165 and renilla luciferase (Rluc). Addition of doxycycline induced Bi-Tet expression of VEGF165 and Rluc significantly compared to baseline (p < 0.05). Expression of VEGF 165 enhanced ES cell proliferation and inhibited apoptosis as determined by Annexin-V staining. For noninvasive imaging, ES cells were transduced with a double fusion (DF) reporter gene consisting of firefly luciferase and enhanced green fluorescence protein (Fluc-eGFP). There was a robust correlation between cell number and Fluc activity (R2 = 0.99). Analysis by immunostaining, histology, and RT-PCR confirmed that expression of Bi-Tet and DF systems did not affect ES cell self-renewal or pluripotency. ES cells were differentiated into beating embryoid bodies expressing cardiac markers such as troponin, Nkx2.5, and β-MHC. Afterward, 5 × 10 5 cells obtained from these beating embryoid bodies or saline were injected into the myocardium of SV129 mice (n = 36) following ligation of the left anterior descending (LAD) artery. Bioluminescence imaging (BLI) and echocardiography showed that VEGF165 induction led to significant improvements in both transplanted cell survival and cardiac function (p < 0.05). This is the first study to demonstrate imaging of embryonic stem cell-mediated gene therapy targeting cardiovascular disease. With further validation, this platform may have broad applications for current basic research and further clinical studies.

Original languageEnglish (US)
Pages (from-to)549-563
Number of pages15
JournalCloning and Stem Cells
Volume9
Issue number4
DOIs
StatePublished - 2007
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Developmental Biology

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