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Construction of genome‐engineered mesenchymal stem cells secreting angiogenic or anti‐inflammatory factors for the treatment of acute kidney injury
Author(s) -
Park HyeJeong,
Cho JeongIn,
Jang HyoJu,
Park EuiJung,
Choi HyoJung,
Kwon TaeHwan
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.649.5
Subject(s) - mesenchymal stem cell , stem cell , cancer research , microbiology and biotechnology , biology
Stem cell therapy has been proposed as a potential therapeutic strategy for acute kidney injury (AKI). By exploiting genome editing and cell sheet technology, we aimed to generate mesenchymal stem cells (MSCs) secreting angiogenic factors [vascular endothelial growth factor (VEGF) and angiopoietin‐1 (ANG1)] or anti‐inflammatory factors [erythropoietin (EPO) and a‐melanocyte stimulating hormone (a‐MSH)] for therapeutic application in AKI. To integrate each gene expression cassette into the safe harbor locus, AAVS1, of the human umbilical cord‐derived MSCs (hUC‐MSCs) chromosome, AAVS1‐targeting Zinc Finger Nuclease (ZFN) or AAVS1‐targeting CRISPR/Cas9 system was exploited. Junction PCR analysis demonstrated the ZFN‐ or CRISPR/Cas9‐aided gene integration in hUC‐MSCs. Flow cytometry and osteogenic and adipogenic differentiation assay revealed that stemness was maintained despite genome engineering. Protein measurement in conditioned media by ELISA and immunoblotting confirmed the production and secretion of each integrated gene product. In vitro assay demonstrated the angiogenic functions (increased HUVEC migration and increased mRNA expression of MMP‐9 and Tie‐2 in co‐cultured HUVEC) of genome‐engineered hUC‐MSCs secreting VEGF or ANG1. For the stem cell therapy in AKI, a scaffold‐free cell sheet system was established using a temperature‐responsive polymer (poly(N‐isopropylacrylamide). Taken together, cell sheet system of hUC‐MSCs secreting angiogenic or anti‐inflammatory factors is successfully established. This is to be examined in animal models of AKI to demonstrate the therapeutic effects of stem cell‐based regenerative strategy against AKI. Support or Funding Information This study was supported by the National Research Foundation of Korea Grants 2014R1A5A2009242 and 2016R1A2B4009365 funded by the Ministry of Science, ICT and Future Planning, Korea This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .