The Assisted Reproductive Technology (ART) Core Laboratory provides investigators with the tools and expertise necessary for incorporating the use of nonhuman primate (NHP) gametes, embryos, follicular cells, and ART-derived techniques into research programs. Additionally, the ART Core is working toward developing additional technologies that will expand its services and increase the utility of NHP models in biomedical research.
ART Core Services
The Core can support or perform the following ARTs:
1. COS and COv Protocol Management. – the ART Core staff can oversee the treatment of investigator animals to ensure injection schedules are maintained and animals are properly managed as well as monitor ovarian activity with routine ultrasound imaging.
- COS: Controlled Ovarian Stimulation is used to produce multiple follicles during a single ovarian cycle
- COv: Controlled Ovulation allows for the managed development of a single, naturally selected follicle
2. Follicle Aspiration and Oocyte Collection – Following either a COv cycle or a stimulated COS cycle, ART Core personnel can track the animal’s ovarian status through circulating steroid (estradiol and progesterone) levels and ultrasound image analysis, coordinate the surgical collection and provide collection medium, isolate oocytes from the follicle aspirates, and carry out in vitro culture of recovered gametes.
3. In Vitro Fertilization – A process whereby semen from investigator animals or ART Core males is incubated in optimized medium with mature oocytes to produce pre-implantation stage embryos. Typically used to evaluate changes in oocyte quality due to treatment effects associated with contraception or infertility based research projects.
4. Intracytoplasmic Sperm Injection (ICSI) – A preferred method for fertilization when the endpoint is embryo production and live offspring. ICSI injects a single spermatozoon into a mature ova using micromanipulation technology.
5. Oocyte/Embryo Micromanipulation – The ART Core can perform several procedures to aid in the study of early embryonic development. This includes injection of oocytes and zygotes with reagents for experimental purposes such as siRNAs/morpholinos for gene knockdown and reagents for gene editing. The Core can also perform trophectoderm biopsies to obtain material for genetic analysis of the embryo
6. Embryo Transfer and Assisted Hatching – Trained ART Core personnel can artificially “hatch” pre-implantation blastocyst stage embryos and surgically transfer them into appropriately staged recipients.
7. Semen Collection and Processing – Freshly collected semen can be used for artificial insemination when the exact timing of insemination is required. Further processing of the semen sample can isolate and determine the number of motile spermatozoa for in vitro fertilization protocols.
8. Embryo and Semen Cryopreservation – In vitro produced embryos can be cryopreserved for storage for later use or analysis. Likewise, semen from genetically valuable males can be collected and stored in the ART Core facility.
9. Media Preparation – All media associated with ART Core protocols are available to researchers and modifications can be made at the investigators request.
10. Experimental Design Assistance – Members of the ART Core team are always happy to answer questions about the resources available to researchers or work together to develop new technology to accomplish research goals.
11. Fetal Sexing –In collaboration with the Hennebold lab, fetal sex may be determined from 2 ml of maternal blood as early as day 20 of gestation.12. Vaginal Swabbing to Confirm Mating –During timed mated breeding, vaginal swabs may be analyzed to determine success during natural breeding attempts.
ART Core Rhesus Macaque Colony
The ART Core maintains a cohort of rhesus macaques from which investigators may request:
1. Oocytes and Follicular Cells – Periovulatory stage oocytes and granulosa cells are collected by laparoscopic follicle aspiration. The timing of the collection can coincide with a specific stage of meiosis in the oocyte. Animals undergo controlled ovary stimulation prior to the collection to maximize the number of possible oocytes recovered.
2. Embryo Transfer – Pre-implantation embryos can be transferred into our recipient animals and monitored for the duration of the pregnancy.
3. Semen Collection and Processing – We maintain males of proven fertility as semen donors from which we can isolate motile spermatozoa for in vitro fertilization, artificial insemination for timed mating protocols, or simple sample analysis.
4. Follicular Fluid Samples – Undiluted follicular fluid can be manually aspirated from specific times during antral follicle development from stimulated or naturally cycling animals.
5. NHP Serum Samples – Characterized banked blood serum samples collected from ART Core NHP females are available to researchers upon request.
6. Ovary Tissue Samples – Beginning Fall 2017, the ART Core will begin banking rhesus ovarian tissue during specific stages of the ovarian cycle. Samples from these ovaries will be made available to investigators as embedded FFPE blocks, frozen in OCT, or as frozen resected tissue.
Integrated Reproductive Services
As part of the Integrated Reproductive Services (IRS), the ART Core works in conjunction with the Timed Mated Breeding program to provide investigators streamlined project support for studies requiring animals of a specific developmental stage, age, or of particular genetics. Services provided by the IRS represent a unique resource for researchers that coordinate projects focused on development of biomedical models for human health and disease. In addition, the IRS provides a single point of contact for budgeting resources and consultation during grant and protocol development and as a conduit for clinicians to addresses reproductive issues for colony management.
Phone number: 503-346-5057Carol Hanna, Ph.D., Director
Cathy Ramsey, B.S., Senior Research Assistant