INTRODUCTION
Human embryonic stem cells (hESCs) have the potential to differentiate into all three germ layers and proliferate in long-term culture in vitro. hESCs can
provide a cell source for the testing of novel therapies, drug screening, and functional genomics applications. Undifferentiated hESCs can be maintained and proliferated on mouse embryonic fibroblasts (MEFs) or human feeder cells. In this protocol, we describe the culture of hESCs in feeder-free conditions on Matrigel with MEF-conditioned medium. This protocol can be used for applications such as genetic modification of hESCs without feeder cell contamination.
RELATED INFORMATION
Protocols for Preparation of Mouse Fibroblast Feeder Cells for Human Embryonic Stem Cell Culture (McElroy and Reijo Pera 2008a), Culturing Human Embryonic Stem Cells with Mouse Embryonic Fibroblast Feeder Cells (McElroy and Reijo Pera 2008b), and Preparation of Human Foreskin Fibroblasts for Human Embryonic Stem Cell Culture (Panula and Reijo Pera 2008) are also available.
Images of undifferentiated hESCs growing in feeder-free conditions are shown in Figure 1 .
Figure 1. (A) A small undifferentiated hESC colony on day 1 after passage on a Matrigel-coated plate. (Black Eye A single hESC colony ready for passage. Images were captured at 50X magnification.