title: The establishment of 20 different human embryonic stem cell lines and subclones; a report on derivation, culture, characterisation and banking.
abstract: This report summarises our efforts in deriving, characterising and banking of 20 different human embryonic stem cell lines. We have derived a large number of human embryonic stem cell lines between 2001 and 2005. One of these cell lines was established under totally xeno-free culture conditions. In addition, several subclones have been established, including a karyoptypical normal clone from a trisomic mother line. A master cell banking system has been utilised in concert with an extensive characterisation programme, ensuring a supply of high quality pluripotent stem cells for further research and development. In this report we also present the first data on a proprietary novel antibody, hES-Cellect, that exhibits high specificity for undifferentiated hES cells. In addition to the traditional manual dissection approach of propagating hES cells, we here also report on the successful approaches of feeder-free cultures as well as single cell cultures based on enzymatic digestion. All culture systems used as reported here have maintained the hES cells in a karyotypical normal and pluripotent state. These systems also have the advantage of being the principal springboards for further scale up of cultures for industrial or clinical applications that would require vastly more cells that can be produced by mechanical means.
authors.affiliation: Cellartis AB Maclagan House, 1 Würzburg Court, Dundee DD2 1FB, UK. mikael.englund@cellartis.com
authors.forename: Mikael C O
authors.initials: MC
authors.lastname: Englund
authors.forename: Gunilla
authors.initials: G
authors.lastname: Caisander
authors.forename: Karin
authors.initials: K
authors.lastname: Noaksson
authors.forename: Katarina
authors.initials: K
authors.lastname: Emanuelsson
authors.forename: Kersti
authors.initials: K
authors.lastname: Lundin
authors.forename: Christina
authors.initials: C
authors.lastname: Bergh
authors.forename: Charles
authors.initials: C
authors.lastname: Hansson
authors.forename: Henrik
authors.initials: H
authors.lastname: Semb
authors.forename: Raimund
authors.initials: R
authors.lastname: Strehl
authors.forename: Johan
authors.initials: J
authors.lastname: Hyllner
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002999:Clone Cells / Q000166:cytology; D003094:Collagen / Q000494:pharmacology; D004210:Dissection; D004338:Drug Combinations; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence; D007621:Karyotyping; D007797:Laminin / Q000494:pharmacology; D051379:Mice; D011509:Proteoglycans / Q000494:pharmacology; D011786:Quality Control; D019098:Telomerase / Q000378:metabolism; D014015:Tissue Banks
chemical_list: D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen; C509186:TERT protein, human; D019098:Telomerase