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  • Generation of induced pluripotent stem cells using skin fibroblasts from patients with myocardial infarction under feeder-free conditions.

Generation of induced pluripotent stem cells using skin fibroblasts from patients with myocardial infarction under feeder-free conditions.

Molecular medicine reports (2014-01-09)
Jun-Quan Li, Ming Cheng, Wei-Chen Tian, Jian Zhang
ABSTRACT

Myocardial infarction (MI) is an increasing medical problem; however, its pathogenesis has yet to be elucidated and more effective treatment strategies are required. Induced pluripotent stem cells (iPSCs) were recently successfully generated using human somatic cells transfected with four transcription factors. The present study aimed to generate iPSCs from cells from patients with myocardial infarction. Six patients who had been diagnosed with myocardial infarction were enrolled in this study. The fibroblast cells from the biopsied skin were reprogrammed using octamer-binding transcription factor 4 (Oct‑4), SRY-related HMG-box gene 2 (Sox‑2), Kruppel-like factor 4 (Klf‑4) and cellular myelocytomatosis oncogene (c‑Myc) transcription factors. The generated cells were identified by karyotyping, in vitro and in vivo differentiation ability and staining for specific markers. These human MI‑iPSCs expressed pluripotent genes and cell surface markers, and exhibited normal proliferation. The iPSCs also showed in vivo and in vitro differentiation ability, as indicated by teratoma and embryoid body formation, respectively. Moreover, the iPSCs differentiated into cardiomyocytes and neuronal cells. In conclusion, human iPSCs were successfully generated from skin fibroblasts from patients with MI under feeder‑independent conditions, which increases their potential suitability for clinical applications. These results may encourage further study of MI pathogenesis and facilitate the development of safe downstream clinical applications of iPSC‑based cell therapies.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Human STEMCCA Constitutive Polycistronic (OKSM) Lentivirus Reprogramming Kit, The Human STEMCCA Constitutive Polycistronic Lentivirus Kit contains high titer polycistronic lentivirus & Polybrene transfection reagent that have been validated for the generation of human iPS cells from human foreskin fibroblasts.
Sigma-Aldrich
Anti-TRA-1-81 Antibody, clone TRA-1-81, clone TRA-1-81, Chemicon®, from mouse