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PNAS 97 (21): 11307-11312

Copyright © 2000 by the National Academy of Sciences.

From the Cover
Developmental Biology
 Effects of eight growth factors on the differentiation of cells derived from human embryonic stem cells

Maya Schuldiner*, Ofra Yanuka*, Joseph Itskovitz-Eldordagger , Douglas A. MeltonDagger ,§, and Nissim Benvenisty*,§

* Department of Genetics, The Institute of Life Sciences, The Hebrew University, Jerusalem 91904, Israel; dagger  Department of Obstetrics and Gynecology, Rambam Medical Center, Faculty of Medicine, The Technion, Haifa 31096, Israel; and Dagger  Department of Molecular and Cellular Biology, Howard Hughes Medical Institute, Harvard University, 7 Divinity Avenue, Cambridge, MA 02138

Contributed by Douglas A. Melton, August 8, 2000

Human embryonic stem (ES) cells are pluripotent cells derived from the inner cell mass of in vitro fertilized human blastocysts. We examined the potential of eight growth factors [basic fibroblast growth factor (bFGF), transforming growth factor beta 1 (TGF-beta 1), activin-A, bone morphogenic protein 4 (BMP-4), hepatocyte growth factor (HGF), epidermal growth factor (EGF), beta  nerve growth factor (beta NGF), and retinoic acid] to direct the differentiation of human ES-derived cells in vitro. We show that human ES cells that have initiated development as aggregates (embryoid bodies) express a receptor for each of these factors, and that their effects are evident by differentiation into cells with different epithelial or mesenchymal morphologies. Differentiation of the cells was assayed by expression of 24 cell-specific molecular markers that cover all embryonic germ layers and 11 different tissues. Each growth factor has a unique effect that may result from directed differentiation and/or cell selection, and we can divide the overall effects of the factors into three categories: growth factors (Activin-A and TGFbeta 1) that mainly induce mesodermal cells; factors (retinoic acid, EGF, BMP-4, and bFGF) that activate ectodermal and mesodermal markers; and factors (NGF and HGF) that allow differentiation into the three embryonic germ layers, including endoderm. None of the growth factors directs differentiation exclusively to one cell type. This analysis sets the stage for directing differentiation of human ES cells in culture and indicates that multiple human cell types may be enriched in vitro by specific factors.

§ To whom reprint requests should be addressed. E-mail: dmelton{at}biohp.harvard.edu or nissimb{at}mail.ls.huji.ac.il.

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