Abstract
Introduction: Embryonic stem cells are pluripotent, thus capable of differentiating into all types of cells derived from the three germ layers. However, the application of embryonic stem cells (ESCs) for preclinical and clinical studies is difficult due to ethical concerns. Induced pluripotent stem cells (iPSCs) are derived from differentiation and have many ESC characteristics. The study herein examines the production of iPSCs from reprogramming of mouse embryonic fibroblasts (MEFs) via transduction with defined factors.
 Methods: MEFs were collected from mouse embryos via a previously published protocol. The cells were transduced with a single polycistronic viral vector encoding mouse cDNAs of Oct3/4, Sox2, Klf4 and c-Myc. Transduced cells were treated and sub-cultured with ESC medium. The cells were evaluated as iPSCs with specific morphology, and expression SSEA-1, Oct3/4, Sox2 and Nanog. In addition, they also were evaluated for pluripotency by assessing alkaline phosphatase (AP) activity and in vivo teratoma formation.
 Results: Under the reprogrammed conditions, the transduced cells displayed a change in morphology, forming ESC-like clusters. These cell clusters strongly expressed pluripotent markers as well as ESC-specific genes. Furthermore, the colonies exhibited higher AP activity and formed teratomas when injected into the murine testis.
 Conclusions: The study herein suggests that MEFs can be reprogrammed into iPSCs using a polycistronic viral vector encoding mouse cDNAs for Oct3/4, Sox2, Klf4 and c-Myc.
Highlights
Embryonic stem cells are pluripotent, capable of differentiating into all types of cells derived from the three germ layers
Embryonic stem cells (ESCs) have the capacity to differentiate into all types of cells derived from the three germ layers (Chambers and Smith, 2004)
Our study aimed to examine the development of induced pluripotent stem cells (iPSCs) via reprogramming of mouse embryonic fibroblasts (MEFs) through transduction with defined factors in polycistronic vector containing 4 pluripotent genes included Oct3/4, Sex determining region Y-box 2 SSEA-1 (Sox2), Klf4 and c-Myc
Summary
Embryonic stem cells are pluripotent, capable of differentiating into all types of cells derived from the three germ layers. Conclusions: The study suggests that MEFs can be reprogrammed into iPSCs using a polycistronic viral vector encoding mouse cDNAs for Oct3/4, Sox, Klf and cMyc. Embryonic stem cells (ESCs) have the capacity to differentiate into all types of cells derived from the three germ layers (Chambers and Smith, 2004). The preferential use of iPSCs can be a strategy that enables the generation of patient-specific stem cells while bypassing the ethical concerns associated with somatic cell nuclear transfer (SCNT) and human embryonic stem cells (Stadtfeld and Hochedlinger, 2010) This platform can be a promising tool for regenerative medicine applications and genetic disease models (Amabile and Meissner, 2009; Hanna et al, 2007; Tateishi et al, 2008; Wernig et al, 2008)
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