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  • Description:
  • Generation of VEGF/hUCB‐MSCs and confirmation of conditional VEGF secretion. (A): Schematic illustration of producing inducible VEGF‐secreting hUCB‐MSC cells and cell sheet transplantation in the rat myocardial infarction model. The human UCB‐MSCs secreting VEGF were seeded onto a cell sheet for transplantation to the myocardial infarction site of rat hearts. (B): The integration of the inducible VEGF secretion DNA cassette into the AAVS1 site following the TALEN‐mediated DNA double‐strand break. HR (L) and HR (R), the left and right arms for homologous recombination. FP and RP, forward primer and reverse primer binding sites for the PCR primers. (C): Junction PCR analysis for the confirmation of the integrated inducible VEGF secretion cassette with two different PCR primer sets. For validation of the gene integration into the UCB‐MSC genome, two sets of primers were used to detect the specific sites; a 2.16 kb fragment with the AAVS1 (forward) and VEGF (reverse) region primers and a 2007 base pair (bp) fragment with thehEF1a (forward) and AAVS1 (reverse) region primers. (D): Western blotting analysis detected inducible VEGF expression in VEGF/hUCB‐MSC cell lysates and (E) inducible VEGF secretion in the VEGF/hUCB‐MSC culture media. The Dox treatment was maintained for 48 hours. (F): Quantification of the secreted VEGF in the VEGF/hUCB‐MSC (Dox+) culture media 2 days after transient transfection and (G) 2 weeks after stable transduction. The amount of VEGF production is indicated as ng per 2 × 106 cells per day on the Y‐axis. The error bars indicate the SEM of three replicate measurements per group. (*p < .05, **p < .01). Abbreviations: AAVS1, adeno‐associated virus integration site 1; Dox, doxycycline; hUCB‐MSCs, human umbilical cord blood‐derived mesenchymal stem cells; HA‐L, left homology arm; HA‐R, right homology arm; HR, homologous recombination; TALEN, transcription activator‐like effector nuclease; VEGF, vascular endothelial growth factor.
  • License:
  • CC-BY-NC-ND
  • Rights Holder:
  • Creative Commons by John Wiley & Sons
  • License Rights Holder:
  • © 2017 AlphaMed Press
  • Asset Type:
  • Image
  • Asset Subtype:
  • Figure
  • Image Orientation:
  • Portrait
  • Image Dimensions:
  • x
  • Image File Size:
  • 388 KB
  • Creator:
  • Hyun‐Min Cho, Pyung‐Hwan Kim, Hyun‐Kyung Chang, Yi‐ming Shen, Kwaku Bonsra, Byung‐Jae Kang, Soo‐Young Yum, Joo‐Hyun Kim, So‐Yeong Lee, Min‐cheol Choi, Hyongbum (Henry) Kim, Goo Jang, Je‐Yoel Cho
  • Credit:
  • Cho, H.-M., Kim, P.-H., Chang, H.-K., Shen, Y.-m., Bonsra, K., Kang, B.-J., Yum, S.-Y., Kim, J.-H., Lee, S.-Y., Choi, M.-c., Kim, H. (., Jang, G., & Cho, J.-Y. (2017). Targeted Genome Engineering to Control VEGF Expression in Human Umbilical Cord Blood‐Derived Mesenchymal Stem Cells: Potential Implications for the Treatment of Myocardial Infarction. STEM CELLS Translational Medicine, 6(3), 1040-1051..
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  • This image was published under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, (CC-BY-NC-ND) which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made. You may purchase a high resolution image file if your request is non-commercial and you are not adapting the image.
  • Property Release:
  • No
  • Model Release:
  • No
  • Purchasable:
  • Yes
  • Sensitive Materials:
  • No
  • Article Authors:
  • Hyun‐Min Cho, Pyung‐Hwan Kim, Hyun‐Kyung Chang, Yi‐ming Shen, Kwaku Bonsra, Byung‐Jae Kang, Soo‐Young Yum, Joo‐Hyun Kim, So‐Yeong Lee, Min‐cheol Choi, Hyongbum (Henry) Kim, Goo Jang, Je‐Yoel Cho
  • Article Copyright Year:
  • 2017
  • Publication Volume:
  • 6
  • Publication Issue:
  • 3
  • Publication Date:
  • 03/01/2017
  • DOI:

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