Fibroblasts derived from type II SMA (GM03813, GM22592 and AIDHC-SP22) and non-SMA (GM03814, AIDHC-NMC1, AIDHC-SC1 and AIDHC-SC2) individuals were grown in DMEM containing 10% EquaFETAL (Atlas Biologicals, Fort Collins, CO), 2 mM L-glutamine (Life Technologies, Grand Island, NY) and 1% penicillin-streptomycin (Life Technologies). GM03813 [30 (link)], GM22592 and GM03814 [30 (link)] fibroblast lines were obtained from Coriell Cell Repositories (Camden, NJ) while the other fibroblast lines were generated at Nemours/Alfred I. duPont Hospital for Children. All type II SMA fibroblast lines used in this study contain 0 copies of SMN1 and 3 copies of SMN2 [31 (link)]. GM03814 fibroblasts [30 (link)] were derived from the carrier mother of GM03813 and contain 1 copy of SMN1 and 5 copies of SMN2 [31 (link)]. The other non-SMA fibroblast lines contain 2 copies of SMN1 and 2 copies of SMN2 [31 (link)]. The fibroblast lines were authenticated using short tandem repeat profiling and digital PCR as described previously [32 (link)].
The mouse motor neuron cell line NSC-34 [33 (link)] and the NSC-34-based reporter lines [18 (link);34 (link)] were maintained in DMEM, 5% EquaFETAL, 2 mM L-glutamine and 1% penicillin/streptomycin. In all instances, the cells were maintained in a humidified chamber at 37°C and 5% CO2.
The mouse motor neuron cell line NSC-34 [33 (link)] and the NSC-34-based reporter lines [18 (link);34 (link)] were maintained in DMEM, 5% EquaFETAL, 2 mM L-glutamine and 1% penicillin/streptomycin. In all instances, the cells were maintained in a humidified chamber at 37°C and 5% CO2.
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