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Arthritis Research and Therapy

, 16:R41

First Online: 04 February 2014Received: 01 May 2013Accepted: 15 January 2014

Abstract

IntroductionSince 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.

MethodsA 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.

ResultsAfter 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.

ConclusionFLSs 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.

AbbreviationsDMEMDulbecco’s modified Eagle’s medium

ESCembryonic stem cell

FBSfetal bovine serum

FLSfibroblast-like synoviocyte

iPSCinduced pluripotent stem cell

OAosteoarthritis

ODMosteogenic differentiation medium

RArheumatoid arthritis

SCIDsevere combined immune deficiency.

Electronic supplementary materialThe online version of this article doi:10.1186-ar4470 contains supplementary material, which is available to authorized users.

Jaecheol Lee, Youngkyun Kim, Hyoju Yi contributed equally to this work.

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Autor: Jaecheol Lee - Youngkyun Kim - Hyoju Yi - Sebastian Diecke - Juryun Kim - Hyerin Jung - Yeri Alice Rim - Seung Min Jung

Fuente: https://link.springer.com/







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