"Use of Human Gametes Obtained from Anonymous Donors for the Production of Human Embryonic Stem-Cell Lines"
by
Susan E. Lanzendorf, Catherine A. Boyd, Diane L. Wright, Suheil Muasher, Sergio Oehninger, and Gary D. Hodgen

[1] Technology Development Center, Jones Institute for Reproductive Medicine, Eastern Virginia Medical School, Norfolk, VA, USA

[2] Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Jones Institute for Reproductive Medicine, Eastern Virginia Medical School, Norfolk, VA, USA

Reprint Requests:
Dr. Susan E. Lanzendorf, Ph.D.
Department of Obstetrics and Gynecology
Jones Institute for Reproductive Medicine
601 Colley Avenue
Norfolk, VA 23507
FAX: 757-446-5262
E-mail: lanzense@evms.edu .

Manuscript received: November 14, 2000
Revised: February 14, 2001
Accepted: February 14, 2001

First presented at the Annual Meeting of the American Society for Reproductive Medicine in San Diego, CA.(October 23-26, 2000).

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 g/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.

Virginia Lab Harvests Human Stem Cells

July 11, 2001; Norfolk, VA (AP) -- Virginia scientists have become the first researchers to create human embryos in the lab for the sole purpose of harvesting their stem cells, according to a study published today. Until now, scientists had derived stem cells only from excess embryos donated from infertility treatments. Other researchers also have derived stem cells from other sources, including fat cells, bone marrow and aborted fetuses. In this case, the scientists approached donors and informed them that their eggs and sperm would be used to develop embryos for stem-cell research. Stem cells can mature into any cell or tissue, and scientists say they may someday be used to repair or replace damaged tissue or organs for disorders such as Alzheimer's, Diabetes, Cancer, Parkinson's Disease, and spinal cord injuries. The work, conducted by researchers at the Jones Institute for Reproductive Medicine in Norfolk appears in the July issue of the journal Fertility and Sterility, the official publication of the American Society of Reproductive Medicine. "The society believes the researchers are the first in the United States to have created embryos explicitly for stem cell research, and it was impressed with their thorough study of the ethics involved, said Society Spokesman Sean Tipton." "At one level, it's cleaner (ethically) than using leftover embryos," Tipton said Wednesday. "There's no question as to what you're going to do with these embryos. You're going to the individuals up front."

President Bush has said he will soon decide whether to allow taxpayer dollars to be used for research on embryonic stem cells. He is under pressure from patient groups that favor the research and opponents who feel the work is inherently unethical.

The Jones Institute work was criticized by religious conservatives opposed to embryo research and from others who have been working to find middle ground in the heated political battle. "I think this is a cautionary tale against starting down the slope," Richard Doerflinger of the National Conference of Catholic Bishops old The Washington Post in Wednesday's Edition. "It's still killing a human being," Mary Petchel, President of the Tidewater Chapter of the Virginia Society for Human Life, told The Virginian-Pilot of Norfolk.

Scientists who conducted the work said several review panels had assessed the ethical implications and concluded that the approach was at least as ethical as using spare frozen embryos. The Institute's Ethics Committee concluded that "the creation of embryos for research purposes was justifiable and that it was our duty to provide humankind with the best understanding of early human development," the team reported in the journal.

The researchers extracted eggs from 12 women, who had signed detailed informed consent documents and were paid $[1,500 - $2,000] each, said William Gibbons, an Eastern Virginia reproductive endocrinologist who was not involved in the work. Of the 162 eggs collected and inseminated by donor sperm, 50 developed into embryos. The researchers destroyed 40 of those to obtain their stem cells, then isolated three cell lines or colonies of stem cells that were maintained in culture. The privately funded study began in 1997 and ended last July.

On the Web:

1. Jones Institute: www.jonesinstitute.org.

2. Fertility journal: asrm.org/Professionals/Fertility&Sterility/fspage.html.