title: Enhanced Reprogramming Efficiency and Kinetics of Induced Pluripotent Stem Cells Derived from Human Duchenne Muscular Dystrophy.
abstract: The generation of disease-specific induced pluripotent stem cells (iPSCs) holds a great promise for understanding disease mechanisms and for drug screening. Recently, patient-derived iPSCs, containing identical genetic anomalies of the patient, have offered a breakthrough approach to studying Duchenne muscular dystrophy (DMD), a fatal disease caused by the mutation in the dystrophin gene. However, development of scalable and high fidelity DMD-iPSCs is hampered by low reprogramming efficiency, the addition of expensive growth factors and slow kinetics of disease-specific fibroblasts. Here, we show an efficient generation of DMD-iPSCs on bFGF secreting human foreskin fibroblast feeders (I-HFF) by employing single polycistronic lentiviral vector for delivering of transcription factors to DMD patient-specific fibroblast cells. Using this method, DMD-iPSCs generated on I-HFF feeders displayed pluripotent characteristics and disease genotype with improved reprogramming efficiency and kinetics over to mouse feeders. Moreover, we were able to maintain disease-specific iPSCs without additional supplementation of bFGF on I-HFF feeders. Our findings offer improvements in the generation of DMD-iPSCs and will facilitate in understanding of pathological mechanisms and screening of safer drugs for clinical intervention.
  
  
  BACKGROUND
  Duchenne Muscular Dystrophy, Reprogramming, Induced pluripotent Stem Cells, Immortalized Human Feeder, Basic Fibroblast Growth Factor, Stem Cell Cassette.
authors.affiliation: Stem Cell Facility, All India Institute of Medical Sciences, N. Delhi, India.
authors.forename: Pooja
authors.initials: P
authors.lastname: Teotia
authors.affiliation: Stem Cell Facility, All India Institute of Medical Sciences, N. Delhi, India.
authors.forename: Sujata
authors.initials: S
authors.lastname: Mohanty
authors.affiliation: Genetics Unit, Department of Pediatrics, All India Institute of Medical Sciences, N. Delhi, India.
authors.forename: Madhulika
authors.initials: M
authors.lastname: Kabra
authors.affiliation: Child Neurology Division, Department of Pediatrics, All India Institute of Medical Sciences, N. Delhi, India.
authors.forename: Sheffali
authors.initials: S
authors.lastname: Gulati
authors.affiliation: Department of Cardio Thoracic Vascular Surgery, All India Institute of Medical Sciences, N. Delhi, India.
authors.forename: Balram
authors.initials: B
authors.lastname: Airan