Derivation, characterization, and differentiation of human embryonic stem cells.

• 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.

Keyword

Animals

Blastocyst

cytology

metabolism

Cell Differentiation

physiology

Cell Line

Cell Lineage

physiology

Coculture Techniques

Embryo

cytology

metabolism

Fibroblasts

cytology

Humans

Immunohistochemistry

Karyotyping

Mice

Pluripotent Stem Cells

cytology

metabolism

Stem Cell Transplantation

Telomerase

metabolism

Transplantation

Heterologous

Publication and Content Type

ref (subject category)

art (subject category)