International Journal of Stem Cells

  • 제16권4호
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  • Pages.438-447
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  • 2023
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  • 2005-3606(pISSN)
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  • 2005-5447(eISSN)
한국줄기세포학회 (Korean Society for Stem Cell Research)
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Assessment of Risks and Benefits of Using Antibiotics Resistance Genes in Mesenchymal Stem Cell-Based Ex-Vivo Therapy

  • Narayan Bashyal (Research Center, CELLeBRAIN, Ltd.) ;
  • Young Jun Lee (Department of Anatomy, Ajou University School of Medicine) ;
  • Jin-Hwa Jung (Research Center, CELLeBRAIN, Ltd.) ;
  • Min Gyeong Kim (Department of Anatomy, Ajou University School of Medicine) ;
  • Kwang-Wook Lee (Department of Anatomy, Ajou University School of Medicine) ;
  • Woo Sup Hwang (Department of Anatomy, Ajou University School of Medicine) ;
  • Sung-Soo Kim (Department of Anatomy, Ajou University School of Medicine) ;
  • Da-Young Chang (Research Center, CELLeBRAIN, Ltd.) ;
  • Haeyoung, Suh-Kim (Research Center, CELLeBRAIN, Ltd.)
  • 투고 : 2023.04.27
  • 심사 : 2023.05.23
  • 발행 : 2023.11.30
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초록

Recently, ex-vivo gene therapy has emerged as a promising approach to enhance the therapeutic potential of mesenchymal stem cells (MSCs) by introducing functional genes in vitro. Here, we explored the need of using selection markers to increase the gene delivery efficiency and evaluated the potential risks associated with their use in the manufacturing process. We used MSCs/CD that carry the cytosine deaminase gene (CD) as a therapeutic gene and a puromycin resistance gene (PuroR) as a selection marker. We evaluated the correlation between the therapeutic efficacy and the purity of therapeutic MSCs/CD by examining their anti-cancer effect on co-cultured U87/GFP cells. To simulate in vivo horizontal transfer of the PuroR gene in vivo, we generated a puromycin-resistant E. coli (E. coli/PuroR) by introducing the PuroR gene and assessed its responsiveness to various antibiotics. We found that the anti-cancer effect of MSCs/CD was directly proportional to their purity, suggesting the crucial role of the PuroR gene in eliminating impure unmodified MSCs and enhancing the purity of MSCs/CD during the manufacturing process. Additionally, we found that clinically available antibiotics were effective in inhibiting the growth of hypothetical microorganism, E. coli/PuroR. In summary, our study highlights the potential benefits of using the PuroR gene as a selection marker to enhance the purity and efficacy of therapeutic cells in MSC-based gene therapy. Furthermore, our study suggests that the potential risk of horizontal transfer of antibiotics resistance genes in vivo can be effectively managed by clinically available antibiotics.

키워드

과제정보

This research was supported by the Korean Fund for Regenerative Medicine (KFRM) grant funded by the Korea government (the Ministry of Science and ICT, the Ministry of Health & Welfare) (21C0715L1), the Korea Drug Development Fund funded by Ministry of Science and ICT, Ministry of Trade, Industry, and Energy, and Ministry of Health and Welfare (RS-2022-00165974, Republic of Korea) and grants from the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare (HI20C0457 to H.S.K.).

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