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We describe a procedure for generating induced pluripotent stem cell lines in rabbits, using retroviral vectors expressing Oct4, Sox2, Klf4, and c-Myc of human origin to reprogram rabbit fibroblasts prepared from an ear skin biopsy. We also provide detailed procedures for characterizing the resulting iPSC lines, including the analysis of pluripotency marker expression by RT-qPCR, immunolabeling, and fluorescent-associated cell sorting, the evaluation of pluripotency by teratoma production and genetic stability by karyotyping.

The laboratory rat (Rattus norvegicus) is now on the leading edge used as a laboratory model system to study pharmacology, toxicology, immunology, nutrition, behavior, and numerous other topics. Therefore, generation of rat induced pluripotent stem cells (iPSCs) through somatic cells reprogramming is a powerful tool for establishing in vitro disease model, development of new protocols for treatment of different diseases, and creating transgenic rat models. Here, we describe a simple adopted protocol for establishing rat iPSCs from different types of somatic cells including rat primary ear fibroblast (PEF) and primary bone marrow cells (BMC).

Varieties of cells orchestrate immune responses. To capture such dynamic phenomena, intravital imaging is an important technique, and it may provide substantial information that is not available using conventional histological analyses. Multiphoton microscopy enables the direct, three-dimensional, and minimally invasive imaging of biological samples with high spatiotemporal resolution, and it has now become the leading method for in vivo imaging studies. Here we describe a basic method for in vivo imaging of dendritic cells (DCs) in the mouse ear skin using multiphoton microscopy.