scholarly journals Hand1-Luc Embryonic Stem Cell Test (Hand1-Luc EST): A novel rapid and highly reproducible in vitro test for embryotoxicity by measuring cytotoxicity and differentiation toxicity using engineered mouse ES cells

2015 ◽  
Vol 40 (2) ◽  
pp. 251-261 ◽  
Author(s):  
Florian Le Coz ◽  
Noriyuki Suzuki ◽  
Hirohisa Nagahori ◽  
Takashi Omori ◽  
Koichi Saito
Keyword(s):  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Es Cells ◽  
Embryonic Stem ◽  
Mouse Es Cells ◽  
Cell Test

scholarly journals Shortening and Improving the Embryonic Stem Cell Test through the Use of Gene Biomarkers of Differentiation

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
Andrea C. Romero ◽  
Eugenio Vilanova ◽  
Miguel A. Sogorb
Keyword(s):  
Gene Expression ◽  
Risk Assessment ◽  
Stem Cell ◽  
Cell Viability ◽  
Embryonic Stem Cell ◽  
Total Duration ◽  
Embryonic Stem ◽  
End Points ◽  
Cell Test

The embryonic Stem cell Test (EST) is a validated assay for testing embryotoxicityin vitro. The total duration of this protocol is 10 days, and its main end-point is based on histological determinations. It is suggested that improvements on EST must be focused toward molecular end-points and, if possible, to reduce the total assay duration. Five days of exposure of D3 cells in monolayers under spontaneous differentiation to 50 ng/mL of the strong embryotoxic 5-fluorouracil or to 75 μg/mL of the weak embryotoxic 5,5-diphenylhydeantoin caused between 20 and 74% of reductions in the expression of the following genes:Pnpla6,Afp,Hdac7,Vegfa, andNes. The exposure to 1 mg/mL of nonembryotoxic saccharin only caused statistically significant reductions in the expression ofNes. These exposures reduced cell viability of D3 cells by 15, 28, and 34%. We applied these records to the mathematical discriminating function of the EST method to find that this approach is able to correctly predict the embryotoxicity of all three above-mentioned chemicals. Therefore, this work proposes the possibility of improve EST by reducing its total duration and by introducing gene expression as biomarker of differentiation, which might be very interesting forin vitrorisk assessment embryotoxicity.

scholarly journals Znanstvena istraživanja u 21. stoljeću - etički aspekti uporabe laboratorijskih životinja i alternativni modeli

2022 ◽  
Vol 53 (5) ◽  
Author(s):  
Ivana Kmetič ◽  
Monika Roller ◽  
Marina Miletić ◽  
Teuta Murati
Keyword(s):  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Embryo Culture ◽  
Embryonic Stem ◽  
Whole Embryo Culture ◽  
Cell Test

U toksikološkim istraživanjima uz uporabu klasičnih (in vivo) istraživanja, primjenjuju se alternativni test sustavi. Korištenje laboratorijskih životinja, embrija, humanog i animalnog tkiva, kultura stanica i fetalnog seruma u istraživanjima smatra se etički problematičnim te se ograničava zakonima, pravilnicima i praksom. Razmatranjem načina kojima bi se neetičnost mogla izbjeći, došlo je do razvoja “3R” načela (akronim za tri pristupa koja bi se trebala provoditi pri istraživanjima na laboratorijskim životinjama), a to su: smanjenje/racionalizacija uporabe laboratorijskih životinja (engl. Reduction), načelo njihove zamjene (engl. Replacement) i poboljšanje uvjeta uzgoja, smještaja i skrbi za životinje (engl. Refinement). Većina je alternativnih testova toksičnosti još uvijek u postupku validacije. Pojedini in vitro testovi za istraživanja embriotoksičnosti (etički posebno osjetljivo područje) koja su priznala nadležna regulatorna tijela, su EST (engl. Embryonic Stem cell Test), WEC (engl. Whole- Embryo Culture) i MM (engl. MicroMass) test. Standardizacija protokola i uvođenje novih in vitro modela predstavlja važan segment napretka u toksikološkim istraživanjima. Znanstvena budućnost tu vidi mogućnost razvoja i implementacije načela etičnosti u istraživanja primjenjujući sustave koji će promišljeno i bez korištenja živih organizama dijelom nadomjestiti metode u biomedicini, veterinarskoj medicini, biotehnologiji i užem smislu - toksikologiji i farmakologiji.

Prevalidation of the Embryonic Stem Cell Test (EST)—A New In Vitro Embryotoxicity Test

1999 ◽  
Vol 13 (4-5) ◽  
pp. 675-681 ◽  
Author(s):  
G Scholz ◽  
E Genschow ◽  
I Pohl ◽  
S Bremer ◽  
M Paparella ◽  
...  
Keyword(s):  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Embryonic Stem ◽  
Cell Test

Validation of the Embryonic Stem Cell Test in the International ECVAM Validation Study on Three In Vitro Embryotoxicity Tests

2004 ◽  
Vol 32 (3) ◽  
pp. 209-244 ◽  
Author(s):  
Elke Genschow ◽  
Horst Spielmann ◽  
Gabriele Scholz ◽  
Ingeborg Pohl ◽  
Andrea Seiler ◽  
...  
Keyword(s):  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Validation Study ◽  
Embryonic Stem ◽  
Cell Test
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