title: Synergistic contribution of SMAD signaling blockade and high localized cell density in the differentiation of neuroectoderm from H9 cells.
abstract: Directed neural differentiation of human embryonic stem cells (ESCs) enables researchers to generate diverse neuronal populations for human neural development study and cell replacement therapy. To realize this potential, it is critical to precisely understand the role of various endogenous and exogenous factors involved in neural differentiation. Cell density, one of the endogenous factors, is involved in the differentiation of human ESCs. Seeding cell density can result in variable terminal cell densities or localized cell densities (LCDs), giving rise to various outcomes of differentiation. Thus, understanding how LCD determines the differentiation potential of human ESCs is important. The aim of this study is to highlight the role of LCD in the differentiation of H9 human ESCs into neuroectoderm (NE), the primordium of the nervous system. We found the initially seeded cells form derived cells with variable LCDs and subsequently affect the NE differentiation. Using a newly established method for the quantitative examination of LCD, we demonstrated that in the presence of induction medium supplemented with or without SMAD signaling blockers, high LCD promotes the differentiation of NE. Moreover, SMAD signaling blockade promotes the differentiation of NE but not non-NE germ layers, which is dependent on high LCDs. Taken together, this study highlights the need to develop innovative strategies or techniques based on LCDs for generating neural progenies from human ESCs.
authors.affiliation: Department of Histology and Embryology, Institute of Stem Cell and Tissue Engineering, Anhui Medical University, Hefei, Anhui 230032, China; Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, CA 94143, USA; Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA. Electronic address: chaol1974@ahmu.edu.cn.
authors.forename: Chao
authors.initials: C
authors.lastname: Liu
authors.affiliation: Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, CA 94143, USA.
authors.forename: Yaping
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Stem Cell Core, Gladstone Institute of Cardiovascular Disease, University of California San Francisco, San Francisco, CA 94158, USA.
authors.forename: Joshua
authors.initials: J
authors.lastname: Arnold
authors.affiliation: Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Bingwei
authors.initials: B
authors.lastname: Lu
authors.affiliation: Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, CA 94143, USA. Electronic address: su.guo@ucsf.edu.
authors.forename: Su
authors.initials: S
authors.lastname: Guo
mesh_terms: D002452:Cell Count / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology*; D006801:Humans; D054258:Neural Plate / Q000166:cytology* / Q000502:physiology*; D015398:Signal Transduction / Q000502:physiology; D051785:Smad Proteins / Q000378:metabolism*
chemical_list: D051785:Smad Proteins
keywords: Differentiation; Human embryonic stem cells; Localized cell density; Neuroectoderm; Seeding cell density