DOI: 10.1126/science.282.5391.1145. Embryonic Stem Cell Lines Derived from Human Blastocysts

Embryonic Stem Cell Lines Derived from Human Blastocysts

10.1126/science.282.5391.1145

Crossref journal-article

American Association for the Advancement of Science (AAAS)

Science (221)

Abstract

Human blastocyst-derived, pluripotent cell lines are described that have normal karyotypes, express high levels of telomerase activity, and express cell surface markers that characterize primate embryonic stem cells but do not characterize other early lineages. After undifferentiated proliferation in vitro for 4 to 5 months, these cells still maintained the developmental potential to form trophoblast and derivatives of all three embryonic germ layers, including gut epithelium (endoderm); cartilage, bone, smooth muscle, and striated muscle (mesoderm); and neural epithelium, embryonic ganglia, and stratified squamous epithelium (ectoderm). These cell lines should be useful in human developmental biology, drug discovery, and transplantation medicine.

Bibliography

Thomson, J. A., Itskovitz-Eldor, J., Shapiro, S. S., Waknitz, M. A., Swiergiel, J. J., Marshall, V. S., & Jones, J. M. (1998). Embryonic Stem Cell Lines Derived from Human Blastocysts. Science, 282(5391), 1145â1147.

Authors 7

James A. Thomson (first)
Joseph Itskovitz-Eldor (additional)
Sander S. Shapiro (additional)
Michelle A. Waknitz (additional)
Jennifer J. Swiergiel (additional)
Vivienne S. Marshall (additional)
Jeffrey M. Jones (additional)

References 27 Referenced 12,562

10.1038/292154a0
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Thomson J. A., Marshall V. S., Curr. Top. Dev. Biol. 38, 133 (1998). (10.1016/S0070-2153(08)60246-X) / Curr. Top. Dev. Biol. by Thomson J. A. (1998)
Thomson J. A., et al., Proc. Natl. Acad. Sci. U.S.A. 92, 7844 (1995). (10.1073/pnas.92.17.7844) / Proc. Natl. Acad. Sci. U.S.A. by Thomson J. A. (1995)
Thirty-six fresh or frozen-thawed donated human embryos produced by IVF were cultured to the blastocyst stage in G1.2 and G2.2 medium (25). Fourteen of the 20 blastocysts that developed were selected for ES cell isolation as described for rhesus monkey ES cells (5). The inner cell masses were isolated by immunosurgery (26) with a rabbit antiserum to BeWO cells and plated on irradiated (35 grays gamma irradiation) mouse embryonic fibroblasts. Culture medium consisted of 80% Dulbecco's modified Eagle's medium (no pyruvate high glucose formulation; Gibco-BRL) supplemented with 20% fetal bovine serum (Hyclone) 1 mM glutamine 0.1 mM β-mercaptoethanol (Sigma) and 1% nonessential amino acid stock (Gibco-BRL). After 9 to 15 days inner cell mass–derived outgrowths were dissociated into clumps either by exposure to Ca 2+ /Mg 2+ -free phosphate-buffered saline with 1 mM EDTA (cell line H1) by exposure to dispase (10 mg/ml; Sigma; cell line H7) or by mechanical dissociation with a micropipette (cell lines H9 H13 and H14) and replated on irradiated mouse embryonic fibroblasts in fresh medium. Individual colonies with a uniform undifferentiated morphology were individually selected by micropipette mechanically dissociated into clumps and replated. Once established and expanded cultures were passaged by exposure to type IV collagenase (1 mg/ml; Gibco-BRL) or by selection of individual colonies by micropipette. Clump sizes of about 50 to 100 cells were optimal. Cell lines were initially karyotyped at passages 2 to 7.
10.1016/0921-8734(91)90018-7
___, Vaziri H., Counter C. M., Allsopp R. C., Exp. Gerontol. 27, 375 (1992). (10.1016/0531-5565(92)90068-B) / Exp. Gerontol. by ___ (1992)
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We thank the personnel of the IVF clinics at the University of Wisconsin School of Medicine and at the Rambam Medical Center for the initial culture and cryopreservation of the embryos used in this study; D. Gardner and M. Lane for the G1.2 and G2.2 media; P. Andrews for the NTERA2 cl.D1 cells and the antibodies used to examine cell surface markers; C. Harris for karyotype analysis; and Geron Corporation for the 293 and MDA cell pellets and for assistance with the telomerase TRAP assay. Supported by the University of Wisconsin (UIR grant 2060) and Geron Corporation (grant 133-BU18).

Dates

Type	When
Created	23 years ago (July 27, 2002, 1:50 a.m.)
Deposited	1 year, 7 months ago (Jan. 12, 2024, 7:13 p.m.)
Indexed	46 minutes ago (Aug. 25, 2025, 11:53 p.m.)
Issued	26 years, 9 months ago (Nov. 6, 1998)
Published	26 years, 9 months ago (Nov. 6, 1998)
Published Print	26 years, 9 months ago (Nov. 6, 1998)

Funders 0

None

BibTeX

@article{Thomson_1998, title={Embryonic Stem Cell Lines Derived from Human Blastocysts}, volume={282}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.282.5391.1145}, DOI={10.1126/science.282.5391.1145}, number={5391}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Thomson, James A. and Itskovitz-Eldor, Joseph and Shapiro, Sander S. and Waknitz, Michelle A. and Swiergiel, Jennifer J. and Marshall, Vivienne S. and Jones, Jeffrey M.}, year={1998}, month=nov, pages={1145–1147} }

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