SBIR-STTR Award

Advances in assisted reproduction would benefit many aspects of medicine and biological research ranging from production of transgenic animals to human infertility treatment. This research aims to develop and prove the worthiness of a microscale instrument that can integrate several in vitro reproductive procedures: oocyte maturation (IVM) fertilization (IVF), and embryo culture (EC). The two main objectives are developing the miniature systems and testing their merit. Many components of successful human IVF procedures today are derived from animal studies, thus all testing will be done with animal oocytes and embryos. Current in vitro systems have stationary medium of constant composition. The proposed micro scale fluidic systems would precisely control the flow of medium past oocytes/embryos tailor the composition of the medium to precisely mimic physiological conditions, and enable automated collection of medium for analysis. A gradually changing medium from a composition for maturation to one for fertilization, to a sequence for embryo culture mimics the slowly changing environment an oocyte experiences in vivo. The precise control of the proposed system avoids the abrupt environmental changes that accompany conventional sequential media processes and should improve IVM. IVF and EC efficiencies. PROPOSED COMMERCIAL APPLICATION: Miniature fluidic systems for in vitro embryo production will reduce costs. may increase reproduction efficiencies and will lead to microscale techniques to analyze preimplantation embryos. The systems will benefit at least three industries: transgenic animal production companies. human IVF clinics. and commercial livestock breeders. The devices are also the forerunners of devices with built-in microanalysis instruments that would have additional impact in basic biological research and future biotech production

An important limitation of current IVP methods is the ability to perform experiments involving large numbers of embryos and repetitions in a highly accurate and repeatable way. Current methods are laborious, inefficient and depend on the skill of the technician. Micro channel embryo culture enables automated high throughput processing because the embryo is not moved, but rather held in a "parking place." This allows the fluid to be exchanged via automated systems rather than attempting to automate embryo manipulation. We propose to develop an automated micro channel embryo culture system to allow rapid optimization of micro channel culture conditions. The system utilizes commercially available automated liquid handling equipment combined with a custom incubation chamber and a digital visualization system. Specific advantages of the proposed automated micro channel culture system (AMCS) include the use of micro channel culture devices (Vitae Cell TM) which facilitates automated processing and allows one to achieve gentle medium changes, a closed system that eliminates perturbations in gas concentrations and temperature, robotically controlled medium changes and visualization eliminating operator variability. The system will initially be used to optimize bovine embryo culture. We propose to investigate parameters such as medium (single vs. sequential), medium replenishment (by flow), gas atmosphere, embryo-to-volume ratio, and medium supplementation in the micro channel system compared to the standard micro-drop.

Thesaurus Terms:
biomedical automation, biomedical equipment development, cow, embryo /fetus culture, growth media egg /ovum, in vitro fertilization, miniature biomedical equipment animal tissue, bioengineering /biomedical engineering