USDOE Human Subjects Research Database, fiscal year 1995

Lawrence Livermore National Laboratory

Project Identifier: LLNL-84-P-101-03

Project Title:

Human Sperm Chromatin and Chromosomes

Principle Investigator: Ms. Laurie A. Gordon

Project started in: 1984

Fiscal Year 1995 Funding for Research on Human Subjects:

Project Funding Information:
Project received funding in Fiscal Year 1995.
Project used human subjects in Fiscal Year 1995.

Funding Sources:

DOE: Office of Health and Environmental Research (OHER)

Amount: $70,000 (Est.)
Comments:

This project is a core function of the Human Genome Center and is funded under A.V. Carrano's Grant. Estimate is based on amount of funding directed for use of this protocol only.

Information on Use of Human Subjects:

Project does not involve use of multiple protocols/subprojects.

IRB Review:
Type of Review: Full Board
Most Recent Approval: May 17, 1995
IRB Approval Number: 84P-101-03

Number of Human Subjects in the Last Reporting Period for this Project: 5
(Reporting periods vary.)

Type of Human Subjects Involvement:

Collection of Bodily Materials:

Collection of personally identifiable bodily materials (blood or blood products, cells, tissue, organs, waste).

• Use of bodily materials collected from subjects specifically for this project.

(a. Objectives, b. Methodology, c. Ionizing Radiation, Radioactive Substances, or Chemical Substances to which human subjects are exposed, d. Involvement of Human Subjects [d.1. procedures used, d.2. risks if any])

The IRB approved "Human Sperm Chromatin and Chromosomes" protocol provides human sperm interphase pronuclei for fluorescence in situ hybridization (FISH) mapping on the Human Genome Project. Sperm-derived pronuclei provide a unique resource of highly decondensed, very extended chromatin for high-resolution ordering of cosmids by FISH and the generation of distance estimates between cosmids. These data are used to construct a metric backbone that greatly facilitates mapping efforts on human chromosome 19.

Sperm donors are drawn from an in-house donor pool (under the direction of Dr. A. Wyrobek, BBRP) or from personal contacts of LLNL employees, e.g. husbands or friends of employees. Confidentiality is maintained by the contact person. Fresh semen samples are collected in a sterile plastic container by donors on a pre-scheduled day and delivered to the lab. The sample is diluted with a special egg-yolk buffer and stored overnight in a refrigerator. The next day the sample is washed, and sperm cells are isolated and co-incubated with oocytes from golden hamsters. After 2-3 hours, the eggs are washed free of sperm and incubated overnight. The following morning they are fixed on microscope slides at stages when the chromatin is in an extended form characteristic of interphase nuclei. Any remaining sperm cells are discarded under regulations pertaining to biohazards.

Sperm pronuclear interphases are hybridized with nick-translated cosmids, which are subsequently detected with fluorescent signals through the microscope (standard FISH techniques). These studies do not involve exposure of human subjects to radiation or chemicals, nor do they involve the establishment of cell cultures, reproductive studies or genetic engineering. There are no known risks in collecting a sperm sample.