Differentiation and Characterization of Dendritic Cells from Human Embryonic Stem Cells

Abstract

Human embryonic stem cells (hESCs) offer great hope in regenerative medicine. Their ability to give rise to almost any type of cell present in the adult body makes them an invaluable tool in finding cures for a variety of diseases. While considerable protocols have been devised to efficiently differentiate hESCs into various cells types including cells of hematopoietic origin, this protocol will focus on the derivation of dendritic cells (DC), a potent antigen-presenting cell. DCs are a highly important arm of the immune system, as they represent one of the few cells that bridge the innate and adaptive systems, leading to effective pathogen clearance. The study of DCs has led to potential applications in diverse fields, such as vaccine development, tumor immunology, and transplantation. In this protocol, we describe two different methods of differentiating hESCs into DCs. The first method uses OP9 bone marrow stromal supporting cells as a coculture system, while the second method utilizes the formation of embryoid body (EB, cellular aggregate) as an approach. To assure the successful outcome and subsequent assessment of the differentiated DCs, supporting protocols have been included in this chapter.

An efficient method for gene silencing in human primary plasmacytoid dendritic cells: silencing of the TLR7/IRF-7 pathway as a proof of concept

“…Moreover, major advances in siRNA modifications and delivery reagents have overcome initial problems of stability and cellular uptake in most cases. However, active cellular nucleases, specific membrane properties and other yet unidentified factors often make siRNA-mediated gene inactivation in human primary cells difficult 19 . Furthermore, some siRNA, also called immunostimulatory siRNA, can be endocytosed and thus trigger TLR7 pathway, leading to nonspecific pDC activation 20 21 .…”

Mesenchymal Stem Cells from Extra-Embryonic Tissues for Tissue Engineering – Regeneration of the Peripheral Nerve

“…Embryonic stem cells (ESCs) were first observed in a pre-implantation embryo by Bongso and colleagues in 1994 [4]. Since then, many cell lines and a multiplicity of tissues have been successfully derived from ESCs and tested in several animal disease models [5][6][7]. Nevertheless, post-transplantation immune-rejection has been a major problem.…”

Assessing the efficacy of mammalian target of rapamycin inhibitors by phosphorylation of p70S6K in CD4-positive cells of liver transplant patients

“…It has been developed for mycophenolate mofetil and cyclosporine A. [8–10] For instance, the determination of inosine-5’-monophosphate dehydrogenase activity was recently suggested to guide mycophenolate mofetil therapy after renal transplantation. This methodology could improve the survival and function of transplant patients.…”