title: Identification of distinct topographical surface microstructures favoring either undifferentiated expansion or differentiation of murine embryonic stem cells.
abstract: The potential of embryonic stem (ES) cells for both self-renewal and differentiation into cells of all three germ layers has generated immense interest in utilizing these cells for tissue engineering or cell-based therapies. However, the ability to culture undifferentiated ES cells without the use of feeder cells as well as means to obtain homogeneous, differentiated cell populations devoid of residual pluripotent ES cells still remain major challenges. Here we have applied murine ES cells to topographically microstructured surface libraries, BioSurface Structure Arrays (BSSA), and investigated whether these could be used to (i) identify topographically microstructured growth supports alleviating the need for feeder cells for expansion of undifferentiated ES cells and (ii) identify specific types of microstructures enforcing differentiation of ES cells. The BSSA surfaces arrays consisted of 504 different topographical microstructures each located in a tester field of 3 x 3 mm. The murine ES cell lines CJ7 and KH2 were seeded upon the BSSA libraries and specific topographical structures facilitating either undifferentiated ES cell growth or enhancing spreading indicative of differentiation of the ES cells were identified. Secondly serial passage of undifferentiated CJ7 ES cells on selected microstructures, identified in the screening of these BSSA libraries, showed that these cells had retained germ-line potential. These results indicate that one specific type of topographical surface microstructures, identified by the BSSA technology, can substitute for feeder cells and that another subset may be used to eliminate undifferentiated ES cells from a population of differentiated ES cells.
authors.affiliation: Interdisciplinary Nanoscience Center (iNANO), Arhus University, Arhus, Denmark.
authors.forename: Lotte D'Andrea
authors.initials: LD
authors.lastname: Markert
authors.forename: Jette
authors.initials: J
authors.lastname: Lovmand
authors.forename: Morten
authors.initials: M
authors.lastname: Foss
authors.forename: Rune Hoff
authors.initials: RH
authors.lastname: Lauridsen
authors.forename: Michael
authors.initials: M
authors.lastname: Lovmand
authors.forename: Ernst-Martin
authors.initials: EM
authors.lastname: Füchtbauer
authors.forename: Annette
authors.initials: A
authors.lastname: Füchtbauer
authors.forename: Karin
authors.initials: K
authors.lastname: Wertz
authors.forename: Flemming
authors.initials: F
authors.lastname: Besenbacher
authors.forename: Finn Skou
authors.initials: FS
authors.lastname: Pedersen
authors.forename: Mogens
authors.initials: M
authors.lastname: Duch
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry; D049109:Cell Proliferation; D002478:Cells, Cultured; D018920:Coculture Techniques; D053595:Embryonic Stem Cells / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology; D051379:Mice; D008856:Microscopy, Fluorescence; D013499:Surface Properties
chemical_list: D000469:Alkaline Phosphatase