title: Derivation of T cells in vitro from mouse embryonic stem cells.
abstract: The OP9/OP9-DL1 co-culture system has become a well-established method for deriving differentiated blood cell types from embryonic and hematopoietic progenitors of both mouse and human origin. It is now used to address a growing variety of complex genetic, cellular and molecular questions related to hematopoiesis, and is at the cutting edge of efforts to translate these basic findings to therapeutic applications. The procedures are straightforward and routinely yield robust results. However, achieving successful hematopoietic differentiation in vitro requires special attention to the details of reagent and cell culture maintenance. Furthermore, the protocol features technique sensitive steps that, while not difficult, take care and practice to master. Here we focus on the procedures for differentiation of T lymphocytes from mouse embryonic stem cells (mESC). We provide a detailed protocol with discussions of the critical steps and parameters that enable reproducibly robust cellular differentiation in vitro. It is in the interest of the field to consider wider adoption of this technology, as it has the potential to reduce animal use, lower the cost and shorten the timelines of both basic and translational experimentation.
authors.affiliation: Department of Biological Sciences, Hunter College and Graduate Center, City University of New York.
authors.forename: Martina
authors.initials: M
authors.lastname: Kučerová-Levisohn
authors.affiliation: Department of Biological Sciences, Hunter College and Graduate Center, City University of New York.
authors.forename: Jordana
authors.initials: J
authors.lastname: Lovett
authors.affiliation: Department of Biological Sciences, Hunter College and Graduate Center, City University of New York.
authors.forename: Armin
authors.initials: A
authors.lastname: Lahiji
authors.affiliation: Sunnybrook Research Institute, Department of Immunology, University of Toronto.
authors.forename: Roxanne
authors.initials: R
authors.lastname: Holmes
authors.affiliation: Sunnybrook Research Institute, Department of Immunology, University of Toronto.
authors.forename: Juan Carlos
authors.initials: JC
authors.lastname: Zúñiga-Pflücker
authors.affiliation: Department of Biological Sciences, Hunter College and Graduate Center, City University of New York; Ortiz@genectr.hunter.cuny.edu.
authors.forename: Benjamin D
authors.initials: BD
authors.lastname: Ortiz
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D018920:Coculture Techniques / Q000379:methods; D003584:Cytological Techniques / Q000379:methods*; D053595:Embryonic Stem Cells / Q000166:cytology*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D051379:Mice; D013601:T-Lymphocytes / Q000166:cytology*