Abstract 2296: Generation of Induced Pluripotent Stem Cells in Healthy Volunteers and Patients With Hereditary Heart Disease
[Background] We recently reported that human induced pluripotent stem (iPS) cells can be induced to differentiate into cardiomyocytes, and characterized their phenotype. Future investigation of human genetic heart disease such hypertrophic cardiomyopathy (HCM) and long QT syndrome, or acquired heart disease such as myocarditis and diabetic cardiomyopathy should be examined using human iPS cell-derived cardiomyocytes. To address this question, we tried to establish various iPS cells which were obtained from healthy volunteers and patients with genetic heart disease.
[Methods and Results]
Human dermal tissue was biopsied (6-mm skin punch biopsy) from healthy volunteers (n=5) and HCM (n=1), and familial atrial and ventricular septal defect (ASD/VSD, n=1), plated onto cell culture dishes, and expanded into dermal fibroblasts.
Fibroblasts were infected with retroviral vectors expressing transcription factors, Oct3/4, Sox2, and Klf4 with or without c-Myc.
Around 30 days after gene transfer, we picked 50 ES cell-like colonies from each donors. Morphological appearance was quite similar to that of ES cells, and did not change after 15 passages.
These iPS cells showed strong expression of Oct3/4, Sox2, Nanog, Rex1, GDF3 and FGF4 in RT-PCR analyses. Immunohistochemistry for Oct3/4, Nanog, SSEA-3, SSEA-4, Tra 1– 60 and Tra 1– 81 also revealed that all established iPS cell lines showed an expression pattern as identical to ES cells.
The HCM patient was a 61 year-old male, who has been refractory to medical therapy, showed severe dyspnea and underwent dual chamber pacing and percutaneous alchohol septal ablation. The iPS cells derived-cardiomyocytes from this patient are now being investigated, and the characteristics were compared with those from healthy volunteers.
From a 30 year-old male with familial ASD/VSD, we also established the patient derived-iPS cells.
[Conclusions] We established iPS cells from five healthy donors, HCM and ASD/VSD patients. Those iPS cells showed typical stem cell morphological appearance and common stem cell markers. Patient specific iPS cells might be a great tool to elucidate the mechanisms of several heart diseases and to lead to the development of new revolutionary therapies.