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- W2006694744 abstract "Amniotic fluid derived mesenchymal stem cells (AF-MSC) have the potential for perinatal therapeutic applications. In addition to their inherent paracrine effect they can also be manipulated to selectively express factors that may have therapeutic value. We investigated the conditions for cell manipulation using integrating lentiviral vectors (LV). Transduction conditions, promotor profiles and selection protocols of AF-MSC are analysed using eGFP as reporter gene. We used redundant AF from genetic amniocentesis samples isolated at 15-19 weeks. AF-MSC were characterized by specific CD and stem cell marker expression and adipo-, osteo- and chondrogenic differentiation assays. Bicistronic LVs expressing eGFP in combination with different selection markers under control of the CMV, CypA, CBA and SFFV promotors were used in the transduction assays. AF-MSCs are easily transduced by LV and addition of polybrene or protamine sulphate did not significantly enhance transduction efficiency. The CMV and SFFV promotors induced the highest expression levels of eGFP. Selection of transduced cells by means of chemotherapeutics and resistance genes (hygromycin and blasticidin) resulted in toxicity and cell death, whereas fluorescent based sorting for eGFP and magnetic sorting using the LV induced tCD34 receptor, resulted in efficient selection. After LV tranduction cells could still be passaged over 15 times and retained their differentiation potential. We demonstrate that AF-MSCs can be easily and effectively transduced using LV and as such be manipulated to overexpress reporter-, selection- and therapeutic genes, which may have perinatal applications." @default.
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- W2006694744 date "2012-01-01" @default.
- W2006694744 modified "2023-09-25" @default.
- W2006694744 title "416: Amniotic fluid derived mesenchymal stem cells as delivery vehicles for therapeutic factors" @default.
- W2006694744 doi "https://doi.org/10.1016/j.ajog.2011.10.434" @default.
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