1ArunA Biomedical, Inc., Athens, Georgia, United States; 2Regenerative Bioscience Center, University of Georgia, Athens, Georgia, USA; 3Axion BioSystems, Atlanta, Georgia, USA

Mouse Pluripotent Stem Cell Motor Neurons Generate Robust Neural Network Activity on Microelectrode Arrays

Steven Stice1,2 , Anirban Majumder1 , Brad Culp1 , Anthony Nicolini3 , Colin Arrowood3, Nicholas Solomotis2, Raymond Swetenburg2

Society of Toxicology (SOT) 2015

Pluripotent stem cell-derived cells that are cost effective and generate robust, uniform results will enhance efforts to identify chemicals that affect the network of activity in the central nervous system (CNS). Currently, rodent neural tissue generates spiking and bursting activity after 7-14 DIV.....

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1Department of Environmental Health Science, University of Georgia, Athens, GA, United States; 2Interdisciplinary Toxicology Program, University of Georgia, Athens, GA, United States; 3Regenerative Bioscience Center, University of Georgia, Athens, GA, United States; 4ORD/NERL/ERD, U.S. EPA, Athens, GA, United States; 5ArunA Biomedical, Inc., Athens, GA, United States

Using Human-Derived Neural Cells As an In Vitro Model for Developmental Neurotoxicity following Exposure to Pesticides

Smith, Mary A.1, 2, 3;Henderson, W. Matthew4; Wallace, Shelley5; Majumder, Anirban5; Amosu, Mayowa1,3; Bian, Xiaoming1; Lu, Kun1, 2, 3;Stice, Steven2, 3, 5

Society of Toxicology (SOT) 2015

The objective of this study was to develop a metabolomics-based DNT assay that includes stages of neural development using progenitor (hNP) and post-mitotic neuronal cells (hN2) to delineate the adverse outcome pathways (AOP) associated with pesticide exposure. Assays were initially validated with known neurotoxic chemicals. In this study, cells were exposed to 0, 0.1, 0.3, 1, 3, 10, 30 and 100 µM of chlorpyrifos, aldicarb, and lindane for 48 hrs. Following exposure, media and cells were separated and biological reactions were quenched prior to extraction, derivatization, and analysis by GC-MS. Metabolomic profiling and subsequent multivariate analysis demonstrated separation for each pesticide class and dose dependent responses were observed at concentrations lower than those eliciting effects in cytotoxicity assays...

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Regenerative Bioscience Center, Interdisciplinary Toxicology Program*, The University of Georgia, Athens, GA 30602, USA

MR Imaging of Human Neural Progenitor Stem Cells: an in vivo Longitudinal Model

Forrest Goodfellow*, Qingying Ming, Xian Wu*, Qun Zhao, Steve Stice*

Society of Toxicology (SOT) 2015

Results: Transplantation and MR tracking of Human Neural Progenitor Cells

  • Colocalization of MRIB (Red) and GFP reporter introduced into the hNP1 cells.
  • Hypointense regions of T2 and T2* weighted anatomy MR images colocalizes MRIB (Red) and GFP in later chick stages (D3 & D6).
  • Clear anatomical contrast in between tissues. (D9 and D11).

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1 Interdisciplinary Toxicology Program, 2 Regenerative Bioscience Center, The University of Georgia, 3 ArunA Biomedical, Inc. Athens, GA, USA.

Bisphenol A Effects on In Vitro Human Neural Development Was Window of Susceptibility Dependent

Xian Wu1,2, Anirban Majumder3, Steven L. Stice1,2,3

Society of Toxicology (SOT) 2015

Multiple central nervous system disorders may be due to extended toxicant exposure during neural development windows of susceptibility (WOS) continuums. Human stem cell can likely model different developmental WOS.
Characterize bisphenol-A (BPA) effects on neural differentiation and cell function (neurite outgrowth) using differentiating human pluripotent stem cell derived neural progenitor during two WOS....

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1ArunA Biomedical Inc., Athens GA, 2Regenerative Bioscience Center, University of Georgia, Athens GA, 3Environmental Health Sciences, University of Georgia, Athens GA

Metabolomics and neurite outgrowth as data rich developmental neurotoxicity assays in a pluripotent stem cell derived human neural model.

Anirban Majumder1, Shelley Wallace1, Xian Wu2, Mayowa Amosu3, Kun Lu3, Mary A. Smith2,3, Steven L. Stice1,2

Society of Toxicology (SOT) 2015

A vast majority of chemical entities, ranging from pesticides to compounds of therapeutic interest, remain untested for potential toxic outcomes on human neural development. Current methods for evaluating developmental neurotoxicity (DNT) rely heavily on animal based testing and non‐uniform cell lines, are often prohibitively expensive, and provide suboptimal predictive value. There is thus a need for rapid, cost effective, in vitro methods to identify such chemicals. To fill this critical gap we evaluated a robust, species representative and physiologically relevant cellular system, combined with high content imaging (HCI) and metabolomics, to address human DNT. Scalable and uniform populations of undifferentiated neural progenitor cells..

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Cell Reprogram. 2013 Apr

Galactosyltransferase knockout pig induced pluripotent stem cells: a cell source for the production of xenotransplant pigs

Liu Y, Yang JY, Lu Y, Yu P, Dove CR, Hutcheson JM, Mumaw JL, Stice SL, West FD

Abstract

The shortage of human organs and tissues for transplant has led to significant interest in xenotransplantation of pig tissues for human patients. However, transplantation of pig organs results in an acute immune rejection, leading to death of the organ within minutes. The α-1,3-galactosyltransferase (GALT) gene has been knocked out in pigs to reduce rejection, yet additional genes need to be modified to ultimately make pig tissue immunocompatible with humans...

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Am J Physiol Cell Physiol. 2013 Feb 15

A novel in vitro model system for smooth muscle differentiation from human embryonic stem cell-derived mesenchymal cells

Guo X, Stice SL, Boyd NL, Chen SY

Abstract

The objective of this study was to develop a novel in vitro model for smooth muscle cell (SMC) differentiation from human embryonic stem cell-derived mesenchymal cells (hES-MCs). We found that hES-MCs were differentiated to SMCs by transforming growth factor-β (TGF-β) in a dose- and time-dependent manner as demonstrated by the expression of SMC-specific genes smooth muscle α-actin, calponin, and smooth muscle myosin heavy chain...

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Stem Cells. 2012 Nov

Neurotrophic effects of leukemia inhibitory factor on neural cells derived from human embryonic stem cells

Majumder A, Banerjee S, Harrill JA, Machacek DW, Mohamad O, Bacanamwo M, Mundy WR, Wei L, Dhara SK, Stice SL

Abstract

Various growth factor cocktails have been used to proliferate and then differentiate human neural progenitor (NP) cells derived from embryonic stem cells (ESC) for in vitro and in vivo studies. However, the cytokine leukemia inhibitory factor (LIF) has been largely overlooked. Here, we demonstrate that LIF significantly enhanced in vitro survival and promoted differentiation of human ESC-derived NP cells. In NP cells, as well as NP-derived neurons, LIF reduced caspase-mediated apoptosis and reduced both spontaneous and H2O2-induced reactive oxygen species in culture. In vitro, NP cell proliferation and the yield of differentiated neurons were significantly higher in the presence of LIF. In NP cells, LIF enhanced cMyc phosphorylation, commonly associated with self-renewal/proliferation...

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Stem Cells Dev. 2012 Sep

Lectins identify glycan biomarkers on glioblastoma-derived cancer stem cells

Tucker-Burden C, Chappa P, Krishnamoorthy M, Gerwe BA, Scharer CD, Heimburg-Molinaro J, Harris W, Usta SN, Eilertson CD, Hadjipanayis CG, Stice SL, Brat DJ, Nash RJ

Abstract

Glioblastoma (GBM) is a highly aggressive primary brain tumor with a poor prognosis. Despite aggressive therapy with surgery, radiotherapy, and chemotherapy, nearly all patients succumb to disease within 2 years. Several studies have supported the presence of stem-like cells in brain tumor cultures that are CD133-positive, are capable of self-renewal, and give rise to all cell types found within the tumor, potentially perpetuating growth. CD133 is a widely accepted marker for glioma-derived cancer stem cells; however, its reliability has been questioned, creating a need for other identifiers of this biologically important subpopulation. We used a panel of 20 lectins to identify differences in glycan expression found in the glycocalyx of undifferentiated glioma-derived stem cells and differentiated cells that arise from them. Fluorescently labeled lectins that specifically recognize α-N-acetylgalactosamine (GalNAc) and α-N-acetylglucosamine (GlcNAc) differentially bound to the cell surface based on the state of cellular differentiation. GalNAc and GlcNAc were highly expressed on the surface of undifferentiated cells and showed markedly reduced expression over a 12-day duration of differentiation...

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Toxicol Sci. 2012 Sep

Metabolomic response of human embryonic stem cell-derived germ-like cells after exposure to steroid hormones

West FD, Henderson WM, Yu P, Yang JY, Stice SL, Smith MA

Abstract

To assess the potential risks of human exposure to endocrine active compounds (EACs), the mechanisms of toxicity must first be identified and characterized. Currently, there are no robust in vitro models for identifying the mechanisms of toxicity in germ cells resulting from EAC exposure. Human embryonic stem cells can differentiate into numerous functional cell types including germ-like cells (GLCs)...

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Stem Cells Dev. 2012 Feb 10

Avian-induced pluripotent stem cells derived using human reprogramming factors

Lu Y, West FD, Jordan BJ, Mumaw JL, Jordan ET, Gallegos-Cardenas A, Beckstead RB, Stice SL

Abstract

To assess the potential risks of human exposure to endocrine active compounds (EACs), the mechanisms of toxicity must first be identified and characterized. Currently, there are no robust in vitro models for identifying the mechanisms of toxicity in germ cells resulting from EAC exposure. Human embryonic stem cells can differentiate into numerous functional cell types including germ-like cells (GLCs)...

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Int J Biomater. 2012;2012:861794

Rapid Heterotrophic Ossification with Cryopreserved Poly(ethylene glycol-) Microencapsulated BMP2-Expressing MSCs

Mumaw J, Jordan ET, Sonnet C, Olabisi RM, Olmsted-Davis EA, Davis AR, Peroni JF, West JL, West F, Lu Y, Stice SL

Abstract

Autologous bone grafting is the most effective treatment for long-bone nonunions, but it poses considerable risks to donors, necessitating the development of alternative therapeutics. Poly(ethylene glycol) (PEG) microencapsulation and BMP2 transgene delivery are being developed together to induce rapid bone formation...

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Stem Cells Dev. 2012 Sep 1

Lectins identify glycan biomarkers on glioblastoma-derived cancer stem cells

Tucker-Burden C, Chappa P, Krishnamoorthy M, Gerwe BA, Scharer CD, Heimburg-Molinaro J, Harris W, Usta SN, Eilertson CD, Hadjipanayis CG, Stice SL, Brat DJ, Nash RJ

Abstract

Glioblastoma (GBM) is a highly aggressive primary brain tumor with a poor prognosis. Despite aggressive therapy with surgery, radiotherapy, and chemotherapy, nearly all patients succumb to disease within 2 years. Several studies have supported the presence of stem-like cells in brain tumor cultures that are CD133-positive, are capable of self-renewal, and give rise to all cell types found within the tumor, potentially perpetuating growth. CD133 is a widely accepted marker for glioma-derived cancer stem cells; however, its reliability has been questioned, creating a need for other identifiers of this biologically important subpopulation....

READ MORE