title: Marmoset induced pluripotent stem cells: Robust neural differentiation following pretreatment with dimethyl sulfoxide.
abstract: The marmoset is an important nonhuman primate model for regenerative medicine. For experimental autologous cell therapy based on induced pluripotent (iPS) cells in the marmoset, cells must be able to undergo robust and reliable directed differentiation that will not require customization for each specific iPS cell clone. When marmoset iPS cells were aggregated in a hanging drop format for 3 days, followed by exposure to dual SMAD inhibitors and retinoic acid in monolayer culture for 3 days, we found substantial variability in the response of different iPS cell clones. However, when clones were pretreated with 0.05-2% dimethyl sulfoxide (DMSO) for 24 hours, all clones showed a very similar maximal response to the directed differentiation scheme. Peak responses were observed at 0.5% DMSO in two clones and at 1% DMSO in a third clone. When patterns of gene expression were examined by microarray analysis, hierarchical clustering showed very similar responses in all 3 clones when they were pretreated with optimal DMSO concentrations. The change in phenotype following exposure to DMSO and the 6 day hanging drop/monolayer treatment was confirmed by immunocytochemistry. Analysis of DNA content in DMSO-exposed cells indicated that it is unlikely that DMSO acts by causing cells to exit from the cell cycle. This approach should be generally valuable in the directed neural differentiation of pluripotent cells for experimental cell therapy.
authors.affiliation: South Texas Veterans Health Care System, San Antonio, TX, United States; Department of Physiology/Barshop Institute, University of Texas Health Science Center at San Antonio, United States.
authors.forename: Zhifang
authors.initials: Z
authors.lastname: Qiu
authors.affiliation: South Texas Veterans Health Care System, San Antonio, TX, United States; Department of Physiology/Barshop Institute, University of Texas Health Science Center at San Antonio, United States.
authors.forename: Anuja
authors.initials: A
authors.lastname: Mishra
authors.affiliation: South Texas Veterans Health Care System, San Antonio, TX, United States; Department of Physiology/Barshop Institute, University of Texas Health Science Center at San Antonio, United States.
authors.forename: Miao
authors.initials: M
authors.lastname: Li
authors.affiliation: South Texas Veterans Health Care System, San Antonio, TX, United States; Department of Physiology/Barshop Institute, University of Texas Health Science Center at San Antonio, United States.
authors.forename: Steven L
authors.initials: SL
authors.lastname: Farnsworth
authors.affiliation: Southwest National Primate Research Center and Texas Biomedical Research Institute, United States.
authors.forename: Bernadette
authors.initials: B
authors.lastname: Guerra
authors.affiliation: Southwest National Primate Research Center and Texas Biomedical Research Institute, United States.
authors.forename: Robert E
authors.initials: RE
authors.lastname: Lanford
authors.affiliation: South Texas Veterans Health Care System, San Antonio, TX, United States; Department of Physiology/Barshop Institute, University of Texas Health Science Center at San Antonio, United States. Electronic address: hornsby@uthscsa.edu.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Hornsby
mesh_terms: D000818:Animals; D002144:Callithrix; D002453:Cell Cycle / Q000187:drug effects / Q000235:genetics; D002454:Cell Differentiation / Q000187:drug effects* / Q000235:genetics; D002460:Cell Line; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D007150:Immunohistochemistry; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results
chemical_list: D012333:RNA, Messenger; D004121:Dimethyl Sulfoxide