Chapter 45 - Genetic Manipulation of Human Embryonic Stem Cells

Abstract

This chapter will present the principle aspects regarding the in vitro genetic manipulation of human ES cells, and the possible uses of the variety of methods currently available. The new technology of establishing human embryonic stem (ES) cell lines has raised hope for breaking new ground in basic and clinical research. Genetic manipulation will be used in applying this technology to biological research, as well as for the specific needs of transplantation medicine. Genetic manipulation has proved to be a key experimental procedure in the field of mouse ES cell research since their initial isolation. Many effective techniques have since been developed for bringing about specific genetic modifications. Some of these methodologies have recently been adapted for the manipulation of human ES cells. These include both transfection and infection protocols, as well as overexpression and homologous recombination procedures. By introduction of genetic modifications to mouse ES cells, a great number of genetically modified animals have been produced, which have proved to be an extremely valuable tool for research. Despite the great potential attributed to human ES cells, much of the basic developmental research using these cells may be limited to in vitro studies. Researchers into human ES cells are forced to adopt developmental models such as embryoid bodies (EBs) for studying early embryogenesis. Possible uses for genetically modified human ES cells could be the creation of cellular models for disease, studying various aspects of early human development, transplantation medicine, and many more.