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It is becoming increasingly imperative to asses the teratogenecity of new chemical
entities (NCEs) and drugs before they enter the market. Ethical limitation
inhibit such studies in pregnant women. The current study attempts to create a
3-Dimentional in vitro platform that mimics human peri-implantation embryonic
development using embryoid bodies (EBs) derived from human embryonic stem
cells (hESCs). A layer of human endometrial cell line (CRL4003) was coated on
layers of extracellular matrix proteins eg; fibronectin and collagen on low melting
agarose (0.5%), to create a bio-mimetic platform similar to the implantation site.
A time course study on EBs was done over a period of 20 days, to understand
the induction of germ lineages. The expression profile of the ectoderm, endoderm,
mesoderm and trophectoderm lineage markers, such as beta III-tubulin, GATA4,
BMP2, Brachury hANP, cTnT, ABCG2, GATA2, BMP4, HAND1 and beta-hCG, were
studied, by SQ and QRT-PCR and immunofluorescence. The lineage composition
of smooth surfaced EBs (SSEBs) at day-6 closely resembled the human peri-
implantation blastocyst. Inhibition in the induction of any of the lineages caused by
exposure to a NCE was confirmed by lack of biomarker expression and by loss of
the ability of the SSEBs to functionally differentiate into particular lineages. Using
proven embryotoxic componds we demonstrated that the model closely mimiced
peri-impantation development and was sensitive even at very low doses. This model
is adoptable to a 96 well plate format for high through put screening of NCEs.
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