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Clonal derivation and characterization of human embryonic stem cell lines.

Artikel i vetenskaplig tidskrift
Författare Nico Heins
Anders Lindahl
Ulrika Karlsson
Marie Rehnström
Gunilla Caisander
Katarina Emanuelsson
Charles Hanson
Henrik Semb
Petter Björquist
Peter Sartipy
Johan Hyllner
Publicerad i Journal of biotechnology
Volym 122
Nummer/häfte 4
Sidor 511-20
ISSN 0168-1656
Publiceringsår 2006
Publicerad vid Institutionen för biomedicin, avdelningen för klinisk kemi och transfusionsmedicin
Sidor 511-20
Språk en
Länkar dx.doi.org/10.1016/j.jbiotec.2005.1...
Ämnesord Biological Markers, Cell Differentiation, Cell Line, cytology, metabolism, Cytogenetic Analysis, Embryo, cytology, Humans, Karyotyping, Pluripotent Stem Cells, cytology, metabolism, Telomerase, metabolism
Ämneskategorier Medicin och Hälsovetenskap

Sammanfattning

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.

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