title: Directed differentiation of embryonic stem cells using a bead-based combinatorial screening method.
abstract: We have developed a rapid, bead-based combinatorial screening method to determine optimal combinations of variables that direct stem cell differentiation to produce known or novel cell types having pre-determined characteristics. Here we describe three experiments comprising stepwise exposure of mouse or human embryonic cells to 10,000 combinations of serum-free differentiation media, through which we discovered multiple novel, efficient and robust protocols to generate a number of specific hematopoietic and neural lineages. We further demonstrate that the technology can be used to optimize existing protocols in order to substitute costly growth factors with bioactive small molecules and/or increase cell yield, and to identify in vitro conditions for the production of rare developmental intermediates such as an embryonic lymphoid progenitor cell that has not previously been reported.
authors.affiliation: Plasticell Ltd, Stevenage Bioscience Catalyst, Stevenage, United Kingdom.
authors.forename: Marina
authors.initials: M
authors.lastname: Tarunina
authors.affiliation: Plasticell Ltd, Stevenage Bioscience Catalyst, Stevenage, United Kingdom; Advanced Centre for Biochemical Engineering, University College London, London, United Kingdom.
authors.forename: Diana
authors.initials: D
authors.lastname: Hernandez
authors.affiliation: Plasticell Ltd, Stevenage Bioscience Catalyst, Stevenage, United Kingdom.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Johnson
authors.affiliation: MRC Centre for Regenerative Medicine/Institute of Stem cell Research, University of Edinburgh, Edinburgh, United Kingdom.
authors.forename: Stanislav
authors.initials: S
authors.lastname: Rybtsov
authors.affiliation: Plasticell Ltd, Stevenage Bioscience Catalyst, Stevenage, United Kingdom.
authors.forename: Vidya
authors.initials: V
authors.lastname: Ramathas
authors.affiliation: Plasticell Ltd, Stevenage Bioscience Catalyst, Stevenage, United Kingdom.
authors.forename: Mylvaganam
authors.initials: M
authors.lastname: Jeyakumar
authors.affiliation: Plasticell Ltd, Stevenage Bioscience Catalyst, Stevenage, United Kingdom.
authors.forename: Thomas
authors.initials: T
authors.lastname: Watson
authors.affiliation: Plasticell Ltd, Stevenage Bioscience Catalyst, Stevenage, United Kingdom.
authors.forename: Lilian
authors.initials: L
authors.lastname: Hook
authors.affiliation: MRC Centre for Regenerative Medicine/Institute of Stem cell Research, University of Edinburgh, Edinburgh, United Kingdom.
authors.forename: Alexander
authors.initials: A
authors.lastname: Medvinsky
authors.affiliation: Advanced Centre for Biochemical Engineering, University College London, London, United Kingdom.
authors.forename: Chris
authors.initials: C
authors.lastname: Mason
authors.affiliation: Plasticell Ltd, Stevenage Bioscience Catalyst, Stevenage, United Kingdom; Progenitor Labs Ltd, Stevenage Bioscience Catalyst, Stevenage, United Kingdom.
authors.forename: Yen
authors.initials: Y
authors.lastname: Choo
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D019070:Cell Lineage; D002478:Cells, Cultured; D020650:Combinatorial Chemistry Techniques / Q000379:methods; D016895:Culture Media, Serum-Free / Q000737:chemistry* / Q000494:pharmacology; D059290:Dopaminergic Neurons / Q000166:cytology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D016130:Immunophenotyping; D054503:Lymphoid Progenitor Cells / Q000378:metabolism; D051379:Mice; D008863:Microspheres; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D010586:Phagocytes / Q000166:cytology / Q000378:metabolism; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
chemical_list: D016895:Culture Media, Serum-Free; D014446:Tyrosine 3-Monooxygenase