Derivation of pluripotent stem cells from cultured human primordial germ cells

Michael J. Shamblott, Joyce Axelman, Shunping Wang, Elizabeth M. Bugg, John W. Littlefield, Peter J. Donovan, Paul D. Blumenthal, George R. Huggins, John D. Gearhart

Research output: Contribution to journalArticle

Abstract

Human pluripotent stem cells would be invaluable for in vitro studies of aspects of human embryogenesis. With the goal of establishing pluripotent stem cell lines, gonadal ridges and mesenteries containing primordial germ cells (PGCs, 5-9 weeks postfertilization) were cultured on mouse STO fibroblast feeder layers in the presence of human recombinant leukemia inhibitory factor, human recombinant basic fibroblast growth factor, and forskolin. Initially, single PGCs in culture were visualized by alkaline phosphatase activity staining. Over a period of 7-21 days, PGCs gave rise to large multicellular colonies resembling those of mouse pluripotent stem cells termed embryonic stem and embryonic germ (EG) cells. Throughout the culture period most cells within the colonies continued to be alkaline phosphatase- positive and tested positive against a panel of five immunological markers (SSEA-1, SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81) that have been used routinely to characterize embryonic stem and EG cells. The cultured cells have been continuously passaged and found to be karyotypically normal and stable. Both XX and XY cell cultures have been obtained. Immunohistochemical analysis of embryoid bodies collected from these cultures revealed a wide variety of differentiated cell types, including derivatives of all three embryonic germ layers. Based on their origin and demonstrated properties, these human PGC-derived cultures meet the criteria for pluripotent stem cells and most closely resemble EG cells.

Original languageEnglish (US)
Pages (from-to)13726-13731
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume95
Issue number23
DOIs
StatePublished - Nov 10 1998

Fingerprint

Pluripotent Stem Cells
Germ Cells
Alkaline Phosphatase
Embryoid Bodies
Feeder Cells
Germ Layers
Mesentery
Colforsin
Embryonic Development
Cultured Cells
Cell Culture Techniques
Fibroblasts
Staining and Labeling
Cell Line
Embryonic Germ Cells

Keywords

  • Alkaline phosphatase
  • Embryoid body
  • Embryonic germ cell
  • Embryonic stem cell

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Shamblott, M. J., Axelman, J., Wang, S., Bugg, E. M., Littlefield, J. W., Donovan, P. J., ... Gearhart, J. D. (1998). Derivation of pluripotent stem cells from cultured human primordial germ cells. Proceedings of the National Academy of Sciences of the United States of America, 95(23), 13726-13731. https://doi.org/10.1073/pnas.95.23.13726

Derivation of pluripotent stem cells from cultured human primordial germ cells. / Shamblott, Michael J.; Axelman, Joyce; Wang, Shunping; Bugg, Elizabeth M.; Littlefield, John W.; Donovan, Peter J.; Blumenthal, Paul D.; Huggins, George R.; Gearhart, John D.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 95, No. 23, 10.11.1998, p. 13726-13731.

Research output: Contribution to journalArticle

Shamblott, MJ, Axelman, J, Wang, S, Bugg, EM, Littlefield, JW, Donovan, PJ, Blumenthal, PD, Huggins, GR & Gearhart, JD 1998, 'Derivation of pluripotent stem cells from cultured human primordial germ cells', Proceedings of the National Academy of Sciences of the United States of America, vol. 95, no. 23, pp. 13726-13731. https://doi.org/10.1073/pnas.95.23.13726
Shamblott, Michael J. ; Axelman, Joyce ; Wang, Shunping ; Bugg, Elizabeth M. ; Littlefield, John W. ; Donovan, Peter J. ; Blumenthal, Paul D. ; Huggins, George R. ; Gearhart, John D. / Derivation of pluripotent stem cells from cultured human primordial germ cells. In: Proceedings of the National Academy of Sciences of the United States of America. 1998 ; Vol. 95, No. 23. pp. 13726-13731.
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