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Open Access Highly Accessed Research article

Generation of disease-specific induced pluripotent stem cells from patients with rheumatoid arthritis and osteoarthritis

Jaecheol Lee123, Youngkyun Kim4, Hyoju Yi4, Sebastian Diecke123, Juryun Kim4, Hyerin Jung4, Yeri Alice Rim4, Seung Min Jung4, Myungshin Kim5, Yong Goo Kim5, Sung-Hwan Park4, Ho-Youn Kim4 and Ji Hyeon Ju1234*

Author Affiliations

1 Division of Cardiology, Department of Medicine, Stanford University School of Medicine, 265 Campus Drive, Room G1120B, Stanford, CA 94305-5454, USA

2 Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, 265 Campus Drive, Room G1120B, Stanford, CA 94305-5454, USA

3 Stanford Cardiovascular Institute, Stanford University School of Medicine, 265 Campus Drive, Room G1120B, Stanford, CA 94305-5454, USA

4 Division of Rheumatology, Department of Internal Medicine, Seoul St. Mary’s Hospital, Institute of Medical Science, College of Medicine, The Catholic University of Korea, #505, Banpo-Dong, Seocho-Gu, Seoul 137-701, South Korea

5 Department of Laboratory Medicine, College of Medicine, Catholic Genetic Laboratory Center, Seoul St. Mary’s Hospital, The Catholic University of Korea, #505, Banpo-Dong, Seocho-Gu, Seoul 137-701, South Korea

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Arthritis Research & Therapy 2014, 16:R41  doi:10.1186/ar4470

Published: 4 February 2014

Abstract

Introduction

Since the concept of reprogramming mature somatic cells to generate induced pluripotent stem cells (iPSCs) was demonstrated in 2006, iPSCs have become a potential substitute for embryonic stem cells (ESCs) given their pluripotency and “stemness” characteristics, which resemble those of ESCs. We investigated to reprogram fibroblast-like synoviocytes (FLSs) from patients with rheumatoid arthritis (RA) and osteoarthritis (OA) to generate iPSCs using a 4-in-1 lentiviral vector system.

Methods

A 4-in-1 lentiviral vector containing Oct4, Sox2, Klf4, and c-Myc was transduced into RA and OA FLSs isolated from the synovia of two RA patients and two OA patients. Immunohistochemical staining and real-time PCR studies were performed to demonstrate the pluripotency of iPSCs. Chromosomal abnormalities were determined based on the karyotype. SCID-beige mice were injected with iPSCs and sacrificed to test for teratoma formation.

Results

After 14 days of transduction using the 4-in-1 lentiviral vector, RA FLSs and OA FLSs were transformed into spherical shapes that resembled embryonic stem cell colonies. Colonies were picked and cultivated on matrigel plates to produce iPSC lines. Real-time PCR of RA and OA iPSCs detected positive markers of pluripotency. Immunohistochemical staining tests with Nanog, Oct4, Sox2, Tra-1-80, Tra-1-60, and SSEA-4 were also positive. Teratomas that comprised three compartments of ectoderm, mesoderm, and endoderm were formed at the injection sites of iPSCs. Established iPSCs were shown to be compatible by karyotyping. Finally, we confirmed that the patient-derived iPSCs were able to differentiate into osteoblast, which was shown by an osteoimage mineralization assay.

Conclusion

FLSs derived from RA and OA could be cell resources for iPSC reprogramming. Disease- and patient-specific iPSCs have the potential to be applied in clinical settings as source materials for molecular diagnosis and regenerative therapy.