Supplementation-dependent differences in the rates of embryonic stem cell self-renewal, differentiation, and apoptosis
Article first published online: 24 SEP 2003
Copyright © 2003 Wiley Periodicals, Inc.
Biotechnology and Bioengineering
Volume 84, Issue 5, pages 505–517, 5 December 2003
How to Cite
Viswanathan, S., Benatar, T., Mileikovsky, M., Lauffenburger, D. A., Nagy, A. and Zandstra, P. W. (2003), Supplementation-dependent differences in the rates of embryonic stem cell self-renewal, differentiation, and apoptosis. Biotechnol. Bioeng., 84: 505–517. doi: 10.1002/bit.10799
- Issue published online: 17 OCT 2003
- Article first published online: 24 SEP 2003
- Manuscript Accepted: 2 JUL 2003
- Manuscript Received: 1 MAY 2003
- National Science Foundation (NSF) Engineering Research Center (ERC); Stem Cell Network, Canada; Whitaker Foundation; Natural Sciences and Engineering Research Council (NSERC) of Canada
- embryonic stem cells;
- cell culture dynamics;
Although it is known that leukemia inhibitory factor (LIF) supports the derivation and expansion of murine embryonic stem (ES) cells, it is unclear whether this is due to inhibitory effects of LIF on ES cell differentiation or stimulatory effects on ES cell survival and proliferation. Using an ES cell line transgenic for green fluorescent protein (GFP) expression under control of the Oct4 promoter, we were able to simultaneously track the responses of live Oct4-GFP-positive (ES) and -negative (differentiated) fractions to LIF, serum, and other growth factors. Our findings show that, in addition to inhibiting differentiation of undifferentiated cells, the administration of LIF resulted in a distinct dose-dependent survival and proliferation advantage, thus enabling the long-term propagation of undifferentiated cells. Competitive responses from the differentiated cell fraction could only be elicited upon addition of serum, fibroblast growth factor-4 (FGF-4), or insulin-like growth factor-1 (IGF-1). The growth factors did not induce additional differentiation of ES cells, but rather they significantly improved the proliferation of already differentiated cells. Our analyses show that, by adjusting culture conditions, including the type and amount of growth factors or cytokines present, the frequency of media exchange, and the presence or absence of serum, we could selectively and specifically alter the survival, proliferation, and differentiation dynamics of the two subpopulations, and thus effectively control population outputs. Our findings therefore have important applications in engineering stem cell culture systems to predictably generate desired stem cells or their derivatives for various regenerative therapies. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng84: 505–517, 2003.