title: Embryonic Stem Cells Cultured in Microfluidic Chambers Take Control of Their Fate by Producing Endogenous Signals Including LIF.
abstract: It is important to understand the role played by endogenous signals in shaping stem cell fate decisions to develop better culture systems and to improve understanding of development processes. In this study, we describe the behavior of mouse embryonic stem cells (mESCs) inside microfluidic chambers (microchambers) operated under conditions of minimal perfusion. mESCs inside microchambers formed colonies and expressed markers of pluripotency in the absence of feeders or pluripotency-inducing signals such as leukemia inhibitory factor (LIF), while mESCs in standard cultureware differentiated rapidly. In a series of experiments, we demonstrate that remarkable differences in stem cell phenotype are due to endogenous production of LIF and other growth factors brought upon by cultivation in confines of a microchamber in the absence of perfusion (dilution). At the protein level, mESCs produced ∼140 times more LIF inside microchambers than under standard culture conditions. In addition, we demonstrate that pluripotent phenotype of stem cells could be degraded by increasing the height (volume) of the microchamber. Furthermore, we show that inhibition of LIF in microchambers, via the JAK/STAT3 pathway, leads to preferential differentiation into mesoderm that is driven by bone morphogenetic protein (BMP)-4. Collectively, we demonstrate for the first time that it is possible to design a cell culture system where stem cell fate is controlled solely by the endogenous signals. Our study may help shift the paradigm of stem cell cultivation away from relying on expensive exogenous molecules such as growth factors and toward designing culture chambers for harnessing endogenous signals. Stem Cells 2016;34:1501-1512.
authors: Department of Biomedical Engineering, University of California, Davis, Davis, California, USA.|Department of Cell and Tissue Biology, University of California San Francisco, San Francisco, California, USA.
authors: Joshua
authors: J
authors: Guild
authors: Department of Biomedical Engineering, University of California, Davis, Davis, California, USA.
authors: Amranul
authors: A
authors: Haque
authors: Department of Biomedical Engineering, University of California, Davis, Davis, California, USA.
authors: Pantea
authors: P
authors: Gheibi
authors: Department of Biomedical Engineering, University of California, Davis, Davis, California, USA.
authors: Yandong
authors: Y
authors: Gao
authors: Department of Biomedical Engineering, University of California, Davis, Davis, California, USA.
authors: Kyung Jin
authors: KJ
authors: Son
authors: Department of Biomedical Engineering, University of California, Davis, Davis, California, USA.
authors: Elena
authors: E
authors: Foster
authors: Department of Cell and Tissue Biology, University of California San Francisco, San Francisco, California, USA.|Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, California, USA.
authors: Sophie
authors: S
authors: Dumont
authors: Department of Biomedical Engineering, University of California, Davis, Davis, California, USA.
authors: Alexander
authors: A
authors: Revzin
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D055415:Bone Morphogenetic Protein 4 / Q000378:metabolism; D019070:Cell Lineage; D000066673:Cell Self Renewal; D002478:Cells, Cultured; D005855:Germ Layers / Q000166:cytology; D053605:Leukemia Inhibitory Factor / Q000378:metabolism*; D008648:Mesoderm / Q000166:cytology; D051379:Mice; D044085:Microfluidics / Q000295:instrumentation*; D008954:Models, Biological; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D015415:Biomarkers; D055415:Bone Morphogenetic Protein 4; D053605:Leukemia Inhibitory Factor
keywords: Differentiation; Embryonic stem cells; Leukemia inhibitory factor; Microfluidics; Pluripotency