Use of human gametes obtained from anonymous donors for the production of human embryonic stem cell lines

Susan E. Lanzendorf, Catherine A. Boyd, Diane L. Wright, Suheil Muasher, Sergio Oehninger, Gary D. Hodgen

Research output: Contribution to journalArticle

Abstract

Objective: To investigate the use of donated gametes for the production of human embryonic stem cell lines. Design: Basic research study. Setting: Assisted Reproductive Technology (ART) program at an academic institution. Patient(s): Consenting oocyte and sperm donors. Intervention(s): None. Main Outcome Measure(s): Oocytes were aspirated from oocyte donors (n = 12) and inseminated with frozen-thawed donor (n = 2) sperm followed by culture of embryos to day 5 or 6 in sequential media. The inner cell masses of expanded blastocysts were isolated using immunosurgery and cultured for 4-11 days on irradiated primary mouse embryonic fibroblasts (PMEFs). Viable cell colonies were passed every 7-10 days onto fresh PMEFs in the presence of leukemia inhibitory factor (0.1 μg/mL) and evaluated for appropriate cell surface markers. Result(s): Immunosurgery of 40 blastocysts resulted in the culture of 18 inner cell masses, which have produced three cell lines. One of these cell lines has been shown to stain positive for alkaline phosphatase and stage-specific embryonic antigen (SSEA)-4 and negative for SSEA-1, express telomerase activity, and produce hCG when allowed to differentiate. Conclusion(s): These findings demonstrate that the future production of human embryonic stem cell lines for therapeutic use is possible with the use of donated gametes. Many ethical issues were considered before the initiation of this study, and it was our goal to ensure that both oocyte and sperm donors understood the nature and purpose of the research before their participating in the study.

Original languageEnglish (US)
Pages (from-to)132-137
Number of pages6
JournalFertility and sterility
Volume76
Issue number1
DOIs
StatePublished - Jul 12 2001

Keywords

  • Blastocysts
  • Embryonic stem cells
  • Gamete donation
  • Inner cell mass

ASJC Scopus subject areas

  • Reproductive Medicine
  • Obstetrics and Gynecology

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    Lanzendorf, S. E., Boyd, C. A., Wright, D. L., Muasher, S., Oehninger, S., & Hodgen, G. D. (2001). Use of human gametes obtained from anonymous donors for the production of human embryonic stem cell lines. Fertility and sterility, 76(1), 132-137. https://doi.org/10.1016/S0015-0282(01)01825-8