Embryotoxicity of lead (II) acetate and aroclor 1254 using a new end point of the embryonic stem cell test.
We developed a new end point of the mouse stem cell test (EST) for developmental neurotoxicity. We tested 2 developmental neurotoxicants, namely, lead (II) acetate and Aroclor 1254, using this EST. Our results showed that lead (II) acetate is nonembryotoxic, and Aroclor 1254 is weakly embryotoxic. To identify a new end point for developmental neurotoxicity, we used the default method of neuronal differentiation for D3 mouse embryonic stem cells with basic fibroblast growth factor (bFGF) and ascorbic acid. Flow cytometry and real-time polymerase chain reaction were used to quantify the inhibition of neuronal differentiation. Our results showed that both lead (II) acetate and Aroclor 1254 reduced the percentage of microtubule-associated protein 2 (MAP-2)-positive cells and the messenger RNA (mRNA) expression level of MAP-2 in a dose-dependent manner. These results suggested that these methods can be used to develop an additional end point of the EST for developmental neurotoxicity using default differentiation of mouse embryonic stem cells.
National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration, Cheongwon-gun, Chungcheongbuk-do, Korea.
Dae Hyun
DH
Baek
Sung Hee
SH
Park
Jae Hyun
JH
Park
Youngju
Y
Choi
Ki Dae
KD
Park
Jin Wook
JW
Kang
Kyoung Suk
KS
Choi
Hyung Soo
HS
Kim
D016475:3T3 Cells; D000818:Animals; D001205:Ascorbic Acid / Q000378:metabolism; D002454:Cell Differentiation; D020111:Chlorodiphenyl (54% Chlorine) / Q000633:toxicity*; D053595:Embryonic Stem Cells / Q000187:drug effects*; D023381:Endpoint Determination; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology; D005434:Flow Cytometry; D015870:Gene Expression; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000187:drug effects; D009942:Organometallic Compounds / Q000633:toxicity*; D012333:RNA, Messenger; D060888:Real-Time Polymerase Chain Reaction; D013723:Teratogens / Q000633:toxicity*
D008869:Microtubule-Associated Proteins; C079240:Mtap2 protein, mouse; D009942:Organometallic Compounds; D012333:RNA, Messenger; D013723:Teratogens; D016222:Fibroblast Growth Factor 2; D020111:Chlorodiphenyl (54% Chlorine); D001205:Ascorbic Acid; C008261:lead acetate