title: An autogeneic feeder cell system that efficiently supports growth of undifferentiated human embryonic stem cells.
abstract: Human embryonic stem cells (hESCs) have great potential as a source of cells for therapeutic uses, but their culture requires the support of mouse or human cells, either directly as a feeder cell layer or indirectly as a source of conditioned medium in feeder-free culture systems. Unfortunately, the risks of cross-transfer of pathogens from xenogeneic or allogeneic feeders or cell by-products limit their medical applications. In addition, not all human feeders support the growth of hESCs equally well, and ethical concerns have been raised regarding the derivation of feeder cells from aborted human fetuses. We report here the culture of hESCs on a novel feeder cell system, comprising fibroblast-like cells derived from the spontaneous differentiation of hESCs. Isogenicity of the hESCs and hESC-derived fibroblasts was confirmed by micro satellite analysis. The nature of the hESC-derived fibroblasts was identified by the expression of specific markers. This feeder system permits continuous growth of undifferentiated and pluripotent hESCs, as demonstrated by the expression of specific hESC markers, by the formation of teratomas after injection of hESCs into severely combined immunodeficient mice, and by in vitro differentiation of hESCs into differentiated cells of ectodermal, endodermal, and mesodermal origin. Feeder cells derived from hESCs offers a potentially more secure autogeneic and genotypically homogenous system for the growth of undifferentiated hESCs.
authors.affiliation: Centre for Stem Cell Biology and Developmental Genetics, University of Newcastle, Central Parkway, Newcastle upon Tyne NE1 3BZ, UK. mio-drag.stojkovic@ncl.ac.uk
authors.forename: Petra
authors.initials: P
authors.lastname: Stojkovic
authors.forename: Majlinda
authors.initials: M
authors.lastname: Lako
authors.forename: Rebecca
authors.initials: R
authors.lastname: Stewart
authors.forename: Stefan
authors.initials: S
authors.lastname: Przyborski
authors.forename: Lyle
authors.initials: L
authors.lastname: Armstrong
authors.forename: Jerome
authors.initials: J
authors.lastname: Evans
authors.forename: Alison
authors.initials: A
authors.lastname: Murdoch
authors.forename: Tom
authors.initials: T
authors.lastname: Strachan
authors.forename: Miodrag
authors.initials: M
authors.lastname: Stojkovic
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D000954:Antigens, Surface / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D049109:Cell Proliferation; D018920:Coculture Techniques / Q000379:methods; D015925:Cryopreservation; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004622:Embryo, Mammalian / Q000166:cytology*; D005347:Fibroblasts / Q000166:cytology*; D005434:Flow Cytometry; D015870:Gene Expression / Q000235:genetics; D006801:Humans; D007621:Karyotyping; D051379:Mice; D016513:Mice, SCID; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D013724:Teratoma / Q000473:pathology; D014157:Transcription Factors / Q000235:genetics
chemical_list: D015703:Antigens, CD; D000954:Antigens, Surface; D017077:Culture Media, Conditioned; D014157:Transcription Factors