Cue, signal, response analysis of murine embryonic stem cell differentiation : multivariable analysis of cytokine and ECM effects on commitment to differentiation

The highly complex nature of developmental cell fate decisions is exemplified by murine embryonic stem cell (ES cell) differentiation, for which almost two decades of study has identified a small set of putative individual molecular cues which are difficult to generalize broadly or discern determinative signals from. Here we offer a combined experimental/computational investigation aimed at elucidating in a systematic manner important synergies between multiple extracellular cues, especially from cytokines and matrix proteins together, and consequent critical intracellular signals within a multi-pathway network governing mouse ES cell differentiation. A factorial design was used to evaluate responses of ES cells to combinations of leukemia inhibitory factor (LIF) and fibroblast growth factor-4 (FGF4) as soluble signals along with fibronectin (Fn) and laminin (Ln) as adhesion substrates. Factorial analysis revealed significant and unexpected cytokine/matrix synergistic effects. These results showed that the outcome elicited by a specific factor depends on the context in which that factor is presented to the cell. For example, LIF is known to completely inhibit the differentiation of ES cells. Its ability to prevent differentiation is unaffected by the presence of FGF4 or Fn. However, when LIF is present with both FGF4 and Fn, increased differentiation is observed. In order to understand underlying mechanistic regulatory causes for these kinds of complicated synergies, we developed a mathematical model capable of decomposing differentiation and proliferation cell fates from overall cell population dynamics. This decomposition showed that the various cue combinations governed one or another, or sometimes multiple, of these cell fates.

Department dc:contributor.department

Massachusetts Institute of Technology. Dept. of Chemical Engineering.

Grantor dc:publisher

Massachusetts Institute of Technology

Year dc:date.issued

2004

Author dc:creator

• Prudhomme, Wendy A. (Wendy Adele), 1975-

Advisor dc:contributor.advisor

• Douglas A. Lauffenburger.

• Chemical Engineering.

Statement dc:rights

• M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission.

Licence dc:rights.uri

• http://dspace.mit.edu/handle/1721.1/7582

Language dc:language.iso

eng

Handle dc:identifier.uri

http://hdl.handle.net/1721.1/30060

OAI identifier oai:identifier

oai:dspace.mit.edu:1721.1/30060