| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
a WiCell Research Institute;
b Wisconsin National Primate Research Center;
c Department of Anatomy, University of Wisconsin-Madison Medical School and The Genome Center of Wisconsin, Madison, Wisconsin, USA
Key Words. Human embryonic stem cell • Fibroblast growth factor
Correspondence: Mark E. Levenstein, Ph.D., WiCell Research Institute, P.O. Box 7365, Madison, Wisconsin 53707-7365, USA. Telephone: 608-262-4568; Fax: 608-262-8474; e-mail: markl{at}wicell.org
Received June 1, 2005;
accepted for publication October 31, 2005.
Human embryonic stem (ES) cells have most commonly been cultured in the presence of basic fibroblast growth factor (FGF2) either on fibroblast feeder layers or in fibroblast-conditioned medium. It has recently been reported that elevated concentrations of FGF2 permit the culture of human ES cells in the absence of fibroblasts or fibroblast-conditioned medium. Herein we compare the ability of unconditioned medium (UM) supplemented with 4, 24, 40, 80, 100, and 250 ng/ml FGF2 to sustain low-density human ES cell cultures through multiple passages. In these stringent culture conditions, 4, 24, and 40 ng/ml FGF2 failed to sustain human ES cells through three passages, but 100 ng/ml sustained human ES cells with an effectiveness comparable to conditioned medium (CM). Two human ES cell lines (H1 and H9) were maintained for up to 164 population doublings (7 and 4 months) in UM supplemented with 100 ng/ml FGF2. After prolonged culture, the cells formed teratomas when injected into severe combined immunodeficient beige mice and expressed markers characteristic of undifferentiated human ES cells. We also demonstrate that FGF2 is degraded more rapidly in UM than in CM, partly explaining the need for higher concentrations of FGF2 in UM. These results further facilitate the large-scale, routine culture of human ES cells and suggest that fibroblasts and fibro-blast-conditioned medium sustain human ES cells in part by stabilizing FGF signaling above a critical threshold.
This article has been cited by other articles:
|
|
 |
|
  M. K. Furue, J. Na, J. P. Jackson, T. Okamoto, M. Jones, D. Baker, R.-I. Hata, H. D. Moore, J. D. Sato, and P. W. Andrews Heparin promotes the growth of human embryonic stem cells in a defined serum-free medium PNAS, September 9, 2008; 105(36): 13409 - 13414. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Yu and J. A. Thomson Pluripotent stem cell lines Genes & Dev., August 1, 2008; 22(15): 1987 - 1997. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. P. Davis, E. S. Ng, M. Costa, A. K. Mossman, K. Sourris, A. G. Elefanty, and E. G. Stanley Targeting a GFP reporter gene to the MIXL1 locus of human embryonic stem cells identifies human primitive streak-like cells and enables isolation of primitive hematopoietic precursors Blood, February 15, 2008; 111(4): 1876 - 1884. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. J. Go, C. Takenaka, and H. Ohgushi Effect of Forced Expression of Basic Fibroblast Growth Factor in Human Bone Marrow-Derived Mesenchymal Stromal Cells J. Biochem., December 1, 2007; 142(6): 741 - 748. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Wang, T. C. Schulz, E. S. Sherrer, D. S. Dauphin, S. Shin, A. M. Nelson, C. B. Ware, M. Zhan, C.-Z. Song, X. Chen, et al. Self-renewal of human embryonic stem cells requires insulin-like growth factor-1 receptor and ERBB2 receptor signaling Blood, December 1, 2007; 110(12): 4111 - 4119. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Naujokat and T. Saric Concise Review: Role and Function of the Ubiquitin-Proteasome System in Mammalian Stem and Progenitor Cells Stem Cells, October 1, 2007; 25(10): 2408 - 2418. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Pick, L. Azzola, A. Mossman, E. G. Stanley, and A. G. Elefanty Differentiation of Human Embryonic Stem Cells in Serum-Free Medium Reveals Distinct Roles for Bone Morphogenetic Protein 4, Vascular Endothelial Growth Factor, Stem Cell Factor, and Fibroblast Growth Factor 2 in Hematopoiesis Stem Cells, September 1, 2007; 25(9): 2206 - 2214. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. L. Boyd, S. K. Dhara, R. Rekaya, E. A. Godbey, K. Hasneen, R. R. Rao, F. D. West III, B. A. Gerwe, and S. L. Stice BMP4 Promotes Formation of Primitive Vascular Networks in Human Embryonic Stem Cell-Derived Embryoid Bodies Experimental Biology and Medicine, June 1, 2007; 232(6): 833 - 843. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Ouyang, R. Ng, and S.-T. Yang Long-Term Culturing of Undifferentiated Embryonic Stem Cells in Conditioned Media and Three-Dimensional Fibrous Matrices Without Extracellular Matrix Coating Stem Cells, February 1, 2007; 25(2): 447 - 454. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Greber, H. Lehrach, and J. Adjaye Fibroblast Growth Factor 2 Modulates Transforming Growth Factor {beta} Signaling in Mouse Embryonic Fibroblasts and Human ESCs (hESCs) to Support hESC Self-Renewal Stem Cells, February 1, 2007; 25(2): 455 - 464. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Babaie, R. Herwig, B. Greber, T. C. Brink, W. Wruck, D. Groth, H. Lehrach, T. Burdon, and J. Adjaye Analysis of Oct4-Dependent Transcriptional Networks Regulating Self-Renewal and Pluripotency in Human Embryonic Stem Cells Stem Cells, February 1, 2007; 25(2): 500 - 510. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Armstrong, O. Hughes, S. Yung, L. Hyslop, R. Stewart, I. Wappler, H. Peters, T. Walter, P. Stojkovic, J. Evans, et al. The role of PI3K/AKT, MAPK/ERK and NF{kappa}{beta} signalling in the maintenance of human embryonic stem cell pluripotency and viability highlighted by transcriptional profiling and functional analysis Hum. Mol. Genet., June 1, 2006; 15(11): 1894 - 1913. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| STEM CELLS | THE ONCOLOGIST | CME | ALPHAMED PRESS JOURNALS |
