Fig. 8
From: Direct reprogramming of urine-derived cells with inducible MyoD for modeling human muscle disease
Overview of options for modeling skeletal muscle disease in cell culture. Primary cell sources for establishing cell cultures include the muscle, skin, urine, and blood. Myoblasts cultured from muscle biopsies require approximately 3 to 4 weeks for expansion and an additional 2 weeks of culture for myotube differentiation [46–48]. Primary dermal fibroblasts isolated from skin biopsies require 3 to 4 weeks for expansion. Fibroblasts can then be transduced with inducible MyoD (iMyoD) lentivirus, expanded, and differentiated to yield myotubes by approximately 8 weeks [12, 14]. Urine cells require 3 to 4 weeks for expansion and a similar time frame for iMyoD-induced myotube formation as dermal fibroblasts. Lymphocytes isolated from blood are typically transformed using Epstein-Barr virus (EBV) to generate lymphoblastoid cell lines (LCLs), which requires 3 to 4 weeks. Fibroblasts, urine-derived cells, and LCLs can all be reprogrammed to induced pluripotent stem cells (iPSCs). Once generated and expanded, iPSCs will be subjected to quality control measures such as immunofluorescence for pluripotency markers, karyotyping, and embryoid body formation or teratoma formation for three germ layers. As each laboratory has its own quality control measures, this process will add variable amount of time for iPSC culture (indicated by hatch marks). iPSCs can be readily differentiated into cardiomyocytes [4, 49, 50]. Improved efficiency for myotube differentiation from iPSCs has been reported, but additional advances are expected to enhance the efficiency of this process [6, 7]