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a Cellartis AB, Göteborg, Sweden;
b Department of Medical Biochemistry, Göteborg University, Göteborg, Sweden;
c Department of Obstetrics and Gynaecology, Sahlgrenska University Hospital, Göteborg, Sweden;
d Institute of Laboratory Medicine, Department of Clinical Chemistry and Transfusion Medicine, Sahlgrenska University Hospital, Göteborg, Sweden
Key Words. Human blastocyst • ICM • Human ES cell • Pluripotency • Subcloning • Differentiation
Henrik Semb, Ph.D., Section of Endocrinology, Lund University, Klinikgatan 26, BMC, B10, SE-22184, Lund, Sweden. Telephone: 46-2223159; Fax: 46-2223600; e-mail: henrik.semb{at}endo.mas.lu.se
The derivation of human embryonic stem (hES) cells establishes a new avenue to approach many issues in human biology and medicine for the first time. To meet the increased demand for characterized hES cell lines, we present the derivation and characterization of six hES cell lines. In addition to the previously described immunosurgery procedure, we were able to propagate the inner cell mass and establish hES cell lines from pronase-treated and hatched blastocysts. The cell lines were extensively characterized by expression analysis of markers characteristic for undifferentiated and differentiated hES cells, karyotyping, telomerase activity measurement, and pluripotency assays in vitro and in vivo. Whereas three of the cell lines expressed all the characteristics of undifferentiated pluripotent hES cells, one cell line carried a chromosome 13 trisomy while maintaining an undifferentiated pluripotent state, and two cell lines, one of which carried a triploid karyotype, exhibited limited pluripotency in vivo. Furthermore, we clonally derived one cell line, which could be propagated in an undifferentiated pluripotent state.
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