| Embryonics Laboratory
Department of Biological Sciences Old Dominion University Norfolk, Virginia 23529 (757)683-5283 Fax Director Dr. R. James Swanson, Ph.D. |
The Embryonics Laboratory at Old Dominion University has been performing mouse
embryo quality control assays for testing human embryo culture conditions since 1980.
Assays have been conducted continuously since that time, making our quality control
assay the longest running assay of its kind in the United States. We offer 2-cell
mouse embryo assays for toxicity evaluation of culture media, transfer media, laboratory
wares, and other products used in all phases of clinical medicine. We have extensive
experience in toxicity testing of gloves, catheters, glues, natural and synthetic rubbers,
metals, and plastics. Assays can be custom tailored to suit your needs. In most cases,
assay results are available by phone or fax within one week of receiving specimens (the
final day of the assay). Hard copies are sent by mail within three days of completion
of assays.
The Embryonics Laboratory also uses a variety of cell lines for co-culturing
with products and substances at listed above. A sample protocol follows:
Mammalian Cell Toxicity Testing
I. Measurement of the additive for pH adjustment as a function of time in medium with and
without protein. [Rationale: These studies inform about the compounds pH value when
"challenged" by conditions encountered in its potential use protocol. Since
mammalian cells require specific minimum essential medium (MEM) requirements for normal
growth, this test series will allow us to choose medium and protein concentrations for
our cell cultures with some prior knowledge of the changing parameters in the test solution.
A. Measurement of pH change over time in various concentrations of MEM medium made
with test medium without protein.
10 x MEM
1.0 x MEM
0.1 x MEM
0.01 x MEM
0.001 x MEM
These data will produce a graph of pH change as a function of time.
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B. Measurement of pH change over time in the medium as made above with the addition
of Bovine Calf Serum (BCS) at the following concentrations
1. 0.1% BSA
2. 0.5% BSA
3. 1.0% BSA
4. 1.0% BCS
5. 5.0% BCS
6. 10.0% BCS
II. Cell inactivation studies [Rationale: These studies inform us about growth
characteristics of mammalian fibroblasts that have been exposed to test medium.]
A. Preparation for initial cells for tests
1. Thaw
2. Grow to one passage
3. Harvest
B. Wash cells in 100% test medium without MEM or a protein source, culture to confluence,
and evaluate cell numbers after culturing and viability before and after culturing.
1. Wash times
a. control, no exposure
b. second exposure
c. 1 minute exposure
d. 2 minute exposure
e. 5 minute exposure
f. 10 minute exposure
g. 30 minute exposure
h. 1 hour exposure
2. Plate one aliquot of each sample, (a) through (h), for culturing and stain one
aliquot of each sample for viability.
3. Trypsinize and collect for evaluation
a. Perform cell count for all samples
b. Stain with trypan blue for viability
C. Repeat item II. B. above with 0.1x MEM/test medium solution exposure