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a Whittier Institute, Department of Pediatrics, University of California, San Diego, California, USA;
b Stem Cell Research, Department of Obstetrics, Gynecology and Reproductive Science, University of California, San Francisco, California, USA
Key Words. Human embryonic stem cells • Pluripotency • Activin A • Defined medium
Correspondence: Alberto Hayek, M.D., Whittier Institute, 9894 Genesee Ave., La Jolla, CA 92037, USA. Telephone: 858-622-7298; Fax: 858-558-3495; e-mail: ahayek{at}ucsd.edu
To date, all human embryonic stem cells (hESCs) available for research require unidentified soluble factors secreted from feeder layers to maintain the undifferentiated state and pluripotency. Activation of STAT3 by leukemia inhibitory factor is required to maintain "stemness" in mouse embryonic stem cells, but not in hESCs, suggesting the existence of alternate signaling pathways for self-renewal and pluripotency in human cells. Here we show that activin A is secreted by mouse embryonic feeder layers (mEFs) and that culture medium enriched with activin A is capable of maintaining hESCs in the undifferentiated state for >20 passages without the need for feeder layers, conditioned medium from mEFs, or STAT3 activation. hESCs retained both normal karyotype and markers of undifferentiated cells, including Oct-4, nanog, and TRA-1-60 and remained pluripotent, as shown by the in vivo formation of teratomas.
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