title: Clonal derivation and characterization of human embryonic stem cell lines.
abstract: Human embryonic stem cells (hESC) are isolated as clusters of cells from the inner cell mass of blastocysts and thus should formally be considered as heterogeneous cell populations. Homogenous hESC cultures can be obtained through subcloning. Here, we report the clonal derivation and characterization of two new hESC lines from the parental cell line SA002 and the previously clonally derived cell line AS034.1, respectively. The hESC line SA002 was recently reported to have an abnormal karyotype (trisomy 13), but within this population of cells we observed rare individual cells with an apparent normal karyotype. At a cloning efficiency of 5%, we established 33 subclones from SA002, out of which one had a diploid karyotype and this subline was designated SA002.5. From AS034.1 we established one reclone designated AS034.1.1 at a cloning efficiency of 0.1%. These two novel sublines express cell surface markers indicative of undifferentiated hESC (SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81), Oct-4, alkaline phosphatase, and they display high telomerase activity. In addition, the cells are pluripotent and form derivatives of all three embryonic germ layers in vitro as well as in vivo. These results, together with the clonal character of SA002.5 and AS034.1.1 make these homogenous cell populations very useful for hESC based applications in drug development and toxicity testing. In addition, the combination of the parental trisomic hESC line SA002 and the diploid subclone SA002.5 provides a unique experimental system to study the molecular mechanisms underlying the pathologies associated with trisomy 13.
authors.affiliation: Cellartis AB, Arvid Wallgrens Backe 20, 413 46 Göteborg, Sweden.
authors.forename: Nico
authors.initials: N
authors.lastname: Heins
authors.forename: Anders
authors.initials: A
authors.lastname: Lindahl
authors.forename: Ulrika
authors.initials: U
authors.lastname: Karlsson
authors.forename: Marie
authors.initials: M
authors.lastname: Rehnström
authors.forename: Gunilla
authors.initials: G
authors.lastname: Caisander
authors.forename: Katarina
authors.initials: K
authors.lastname: Emanuelsson
authors.forename: Charles
authors.initials: C
authors.lastname: Hanson
authors.forename: Henrik
authors.initials: H
authors.lastname: Semb
authors.forename: Petter
authors.initials: P
authors.lastname: Björquist
authors.forename: Peter
authors.initials: P
authors.lastname: Sartipy
authors.forename: Johan
authors.initials: J
authors.lastname: Hyllner
mesh_terms: D015415:Biomarkers; D002454:Cell Differentiation; D002460:Cell Line / Q000166:cytology / Q000378:metabolism*; D020732:Cytogenetic Analysis; D004622:Embryo, Mammalian / Q000166:cytology*; D006801:Humans; D066298:In Vitro Techniques; D007621:Karyotyping; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D019098:Telomerase / Q000378:metabolism
chemical_list: D015415:Biomarkers; D019098:Telomerase