Developing osteoblasts as an endpoint for the mouse embryonic stem cell test.
The mouse Embryonic Stem cell Test (EST) using cardiomyocyte differentiation is a promising in vitro assay for detecting potential embryotoxicity; however, the addition of another differentiation endpoint, such as osteoblasts, may improve the predictive value of the test. A number of variables such as culture conditions and starting cell number were investigated. A 14 day direct plating method of D3 mouse embryonic stem cells (mESCs) was used to test the predictivity of osteoblast differentiation as an endpoint in the EST. Twelve compounds were tested using the prediction model developed in the ECVAM validation study. Eight of the compounds selected from the EST validation study served as model compounds; four additional compounds known to produce skeletal defects were also tested. Our results indicate comparable chemical classification between the validated cardiomyocyte endpoint and the osteoblast endpoint. These results suggest that differentiation to osteoblasts may provide confirmatory information in predicting embryotoxicity.
Division of Systems Biology, National Center for Toxicological Research, Food and Drug Administration. Electronic address: Xinrong.chen@fda.hhs.gov.
Xinrong
X
Chen
Division of Systems Biology, National Center for Toxicological Research, Food and Drug Administration. Electronic address: Deborah.Hansen@fda.hhs.gov.
Deborah K
DK
Hansen
Division of Systems Biology, National Center for Toxicological Research, Food and Drug Administration. Electronic address: Gwenn.Merry@fda.hhs.gov.
Gwenn
G
Merry
Lee University. Electronic address: ChristiandeJarnette@comcast.net.
Christian
C
DeJarnette
Division of Systems Biology, National Center for Toxicological Research, Food and Drug Administration. Electronic address: Vivian_nolen@att.net.
Greg
G
Nolen
Division of Systems Biology, National Center for Toxicological Research, Food and Drug Administration. Electronic address: Daniel.Sloper@fda.hhs.gov.
Daniel
D
Sloper
Center for Drug Evaluation and Research, Food and Drug Administration. Electronic address: Jedward.fisher@fda.hhs.gov.
J Edward
JE
Fisher
Center for Drug Evaluation and Research, Food and Drug Administration. Electronic address: Wafa.Harrouk@fda.hhs.gov.
Wafa
W
Harrouk
Center for Drug Evaluation and Research, Food and Drug Administration. Electronic address: Melissa.Tassinari@fda.hhs.gov.
Melissa S
MS
Tassinari
Division of Systems Biology, National Center for Toxicological Research, Food and Drug Administration. Electronic address: Amy.Inselman@fda.hhs.gov.
Amy L
AL
Inselman
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D056945:Hep G2 Cells; D006801:Humans; D051379:Mice; D000066450:Mouse Embryonic Stem Cells; D032383:Myocytes, Cardiac / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D015203:Reproducibility of Results; D013723:Teratogens / Q000633:toxicity*
D013723:Teratogens; D000469:Alkaline Phosphatase; D002118:Calcium
Alternative model; Cardiomyocyte; Embryonic stem cell test; Embryotoxicity; In vitro; Osteoblast