Chapter 4 —— 72 —— derived primary human mesenchymal stem cells (hMSCs) were kept in Minimum Essential Medium α (MEM-α; Thermo Fisher Scientific; cat. no.: 22561–021) with 10% FBS, 100 U ml-1 penicillin/streptomycin (Thermo Fisher Scientific; cat. no.: 15140–122), 1 × non-essential amino acids (Thermo Fisher Scientific; cat. no.: 11140–050), 1 × GlutaMax supplement (Thermo Fisher Scientific; cat. no.: 35050–061) and 5 ng ml-1 Recombinant Human Fibroblast Growth Factor-basic (FGF-2; Peprotech; cat. no.: 100– 18B), at 37℃ in a 5% CO2 atmosphere. The collection of human primary cells from bone marrow was, in accordance with the Best Practices Code of the Dutch Federation of Biomedical Scientific Societies, carried out from anonymous ‘left-over’ surgery material. No informed consent was demanded for the usage of such anonymous and non-traceable body materials and the institutional ethics committee of the LUMC waived the permission from the donor. The human muscle progenitor myoblasts, a gift from Vincent Mouly (Sorbonne University, Paris, France), were maintained at 37℃ in a 5% CO2 atmosphere in Ham’s F-10 Nutrient Mix containing 1 × GlutaMAX supplement (Thermo Fisher Scientific; cat. no.: 41550–021), 20% heatinactivated FBS (HI FBS; Thermo Fisher Scientific; cat. no.: 10500–064), 10 ng ml-1 FGF-2, 1 μM Dexamethasone (Sigma-Aldrich; cat. no.: D2915) and 100 U ml-1 penicillin/streptomycin. Recombinant DNA The generation of all the plasmids used herein followed standard recombinant DNA techniques. The annotated maps and nucleotide sequences of the constructs used for the assembly of the nuclease-encoding high-capacity AdVP.C9KARA and AdVP.eC94NLSGS1 particles (i.e. U67_pHCAdVP.C9KARA and S90_pHC-AdVP.eC94NLSGS1, respectively); for the assembly of the single-stranded AAV donor vectors without gRNA expression units AAV-HRS1, AAV-HMEJS1, AAV-HRL.S1 and AAVHMEJL.S1 (i.e. BG14_pAAV-HRS1, BG13_pAAV-HMEJS1, BE22_pAAVHRL.S1 and AG68_pAAV-HMEJL.S1, respectively), for the assembly of the single-stranded AAV donor vectors with gRNA expression units AAV-
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