Treatment of human embryos with the TGF  inhibitor SB431542 increases epiblast proliferation and permits successful human embryonic stem cell derivation

STUDY QUESTION Is there an effect of the TGF beta inhibitor SB431542 (SB) on the epiblast compartment of human blastocysts, and does it affect subsequent human embryonic stem cell (hESC) derivation? SUMMARY ANSWER SB increases the mean number of NANOG-positive cells in the inner cell mass (ICM), and...

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Published in:Human reproduction (Oxford) 2014-01, Vol.29 (1), p.41-48
Main Authors: Van der Jeught, M., Heindryckx, B., O'Leary, T., Duggal, G., Ghimire, S., Lierman, S., Van Roy, N., Chuva de Sousa Lopes, S. M., Deroo, T., Deforce, D., De Sutter, P.
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Language:English
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Activin
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Summary:STUDY QUESTION Is there an effect of the TGF beta inhibitor SB431542 (SB) on the epiblast compartment of human blastocysts, and does it affect subsequent human embryonic stem cell (hESC) derivation? SUMMARY ANSWER SB increases the mean number of NANOG-positive cells in the inner cell mass (ICM), and allows for subsequent hESC derivation. WHAT IS KNOWN ALREADY It is known that inhibition of TGF beta by SB has a positive effect on mouse ESC self-renewal, while active TGF beta signalling is needed for self-renewal of primed ESC. STUDY DESIGN, SIZE, DURATION From December 2011 until March 2012, 263 donated spare embryos were used from patients who had undergone IVF/ICSI in our centre. PARTICIPANTS/MATERIALS, SETTING, METHODS Donated human embryos were cultured in the presence of SB or Activin A, and immunocytochemistry was performed on Day 6 blastocysts for NANOG and GATA6. Moreover, blastocysts were used for the derivation of hESC, with or without exposure to SB. MAIN RESULTS AND THE ROLE OF CHANCE Immunocytochemistry revealed a significantly higher number of NANOG-positive ICM cells in the SB group compared with the control (12.0 plus or minus 5.9 versus 6.1 plus or minus 4.7), while no difference was observed in the Activin A group compared with other groups (6.7 plus or minus 3.7). The number of GATA6-positive ICM cells did not differ between the SB, Activin A and control group (8.8 plus or minus 4.3, 8.0 plus or minus 4.6 and 7.2 plus or minus 4.0, respectively). Blocking TGF beta signalling did not prevent subsequent hESC line derivation. LIMITATIONS, REASONS FOR CAUTION The number of human blastocysts available for this study was too low to reveal if the observed increase in NANOG-positive epiblast cells after exposure to SB affected the efficiency of hESC derivation (12.5% compared with 16.7%). WIDER IMPLICATIONS OF THE FINDINGS This work can contribute to the derivation of naive hESC lines in the future. STUDY FUNDING/COMPETING INTEREST(S) M.V.d.J. is holder of a Ph.D. grant of the Agency for Innovation by Science and Technology (IWT, grant number SB093128), Belgium. G.D. and this research are supported by the Research Foundation Flanders (FWO), grant number FWO-3G062910) and a Concerted Research Actions funding from BOF (Bijzonder Onderzoeksfonds University Ghent, grant number BOF GOA 01G01112). S.M.C.d. S.L. is supported by the Netherlands Organization of Scientific Research (NWO) (ASPASIA 015.007.037) and the Interuniversity Attraction Poles (PAI)
ISSN:0268-1161
1460-2350
DOI:10.1093/humrep/det400