Advanced search
1 file | 1.02 MB Add to list

Derivation of human embryonic stem cells using a post-inner cell mass intermediate

(2013) NATURE PROTOCOLS. 8(2). p.254-264
Author
Organization
Abstract
Little is known about the true developmental origin of human embryonic stem cells (hESCESCESCs) or the events that initiate their generation. Recently, we have shown that hESCESCESCs originate from a post–inner cell mass (ICM) intermediate (PICMI), a unique transient epiblast-like structure that is different from both its ICM progenitor and its subsequent hESCESCESC fate. As a closer progenitor of hESCESCESCs than the ICM, the PICMI could be used to provide further insight into the human pluripotent state. Here we provide a detailed (7-d) protocol for the culture of the human preimplantation embryos in order to derive the PICMI. Subsequent identification and cryopreservation of the PICMI are described, in addition to hESCESCESC derivation. The initial hESCESCESC outgrowth is visible within 2–7 d after PICMI plating. By using the protocol provided, we observed PICMI formation in 21.3% of plated blastocysts with good-quality ICMs. Of the PICMIs used for hESCESCESC derivation, 80.6% showed hESCESCESC outgrowth after further culture.
Keywords
FEEDER CELLS, IN-VITRO FERTILIZATION, LINES, SCORE, ESTABLISHMENT, BLASTOCYST, DIFFERENTIATION, PLURIPOTENCY, IMPLANTATION, FIBROBLASTS

Downloads

  • (...).pdf
    • full text
    • |
    • UGent only
    • |
    • PDF
    • |
    • 1.02 MB

Citation

Please use this url to cite or link to this publication:

MLA
O’Leary, Thomas, et al. “Derivation of Human Embryonic Stem Cells Using a Post-Inner Cell Mass Intermediate.” NATURE PROTOCOLS, vol. 8, no. 2, 2013, pp. 254–64, doi:10.1038/nprot.2012.157.
APA
O’Leary, T., Heindryckx, B., Lierman, S., Van der Jeught, M., Duggal, G., De Sutter, P., & Lopes, S. M. C. de S. (2013). Derivation of human embryonic stem cells using a post-inner cell mass intermediate. NATURE PROTOCOLS, 8(2), 254–264. https://doi.org/10.1038/nprot.2012.157
Chicago author-date
O’Leary, Thomas, Björn Heindryckx, Sylvie Lierman, Margot Van der Jeught, Galbha Duggal, Petra De Sutter, and Susana M Chuva de Sousa Lopes. 2013. “Derivation of Human Embryonic Stem Cells Using a Post-Inner Cell Mass Intermediate.” NATURE PROTOCOLS 8 (2): 254–64. https://doi.org/10.1038/nprot.2012.157.
Chicago author-date (all authors)
O’Leary, Thomas, Björn Heindryckx, Sylvie Lierman, Margot Van der Jeught, Galbha Duggal, Petra De Sutter, and Susana M Chuva de Sousa Lopes. 2013. “Derivation of Human Embryonic Stem Cells Using a Post-Inner Cell Mass Intermediate.” NATURE PROTOCOLS 8 (2): 254–264. doi:10.1038/nprot.2012.157.
Vancouver
1.
O’Leary T, Heindryckx B, Lierman S, Van der Jeught M, Duggal G, De Sutter P, et al. Derivation of human embryonic stem cells using a post-inner cell mass intermediate. NATURE PROTOCOLS. 2013;8(2):254–64.
IEEE
[1]
T. O’Leary et al., “Derivation of human embryonic stem cells using a post-inner cell mass intermediate,” NATURE PROTOCOLS, vol. 8, no. 2, pp. 254–264, 2013.
@article{3237101,
  abstract     = {{Little is known about the true developmental origin of human embryonic stem cells (hESCESCESCs) or the events that initiate their generation. Recently, we have shown that hESCESCESCs originate from a post–inner cell mass (ICM) intermediate (PICMI), a unique transient epiblast-like structure that is different from both its ICM progenitor and its subsequent hESCESCESC fate. As a closer progenitor of hESCESCESCs than the ICM, the PICMI could be used to provide further insight into the human pluripotent state. Here we provide a detailed (7-d) protocol for the culture of the human preimplantation embryos in order to derive the PICMI. Subsequent identification and cryopreservation of the PICMI are described, in addition to hESCESCESC derivation. The initial hESCESCESC outgrowth is visible within 2–7 d after PICMI plating. By using the protocol provided, we observed PICMI formation in 21.3% of plated blastocysts with good-quality ICMs. Of the PICMIs used for hESCESCESC derivation, 80.6% showed hESCESCESC outgrowth after further culture.}},
  author       = {{O'Leary, Thomas and Heindryckx, Björn and Lierman, Sylvie and Van der Jeught, Margot and Duggal, Galbha and De Sutter, Petra and Lopes, Susana M Chuva de Sousa}},
  issn         = {{1754-2189}},
  journal      = {{NATURE PROTOCOLS}},
  keywords     = {{FEEDER CELLS,IN-VITRO FERTILIZATION,LINES,SCORE,ESTABLISHMENT,BLASTOCYST,DIFFERENTIATION,PLURIPOTENCY,IMPLANTATION,FIBROBLASTS}},
  language     = {{eng}},
  number       = {{2}},
  pages        = {{254--264}},
  title        = {{Derivation of human embryonic stem cells using a post-inner cell mass intermediate}},
  url          = {{http://dx.doi.org/10.1038/nprot.2012.157}},
  volume       = {{8}},
  year         = {{2013}},
}

Altmetric
View in Altmetric
Web of Science
Times cited: