Culturing in vitro produced blastocysts in sequential media promotes ES cell derivation

Jun Liu, Luc Schoonjans, S Tielens, Frank Speleman, Maria Cornelissen, P de Sutter, M Dhont, J Van Der Elst

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10 Citations (Scopus)


Embryonic stem (ES) cell lines are routinely derived from in vivo produced blastocysts. We investigated the efficiency of ES cells derivation from in vitro produced blastocysts either in monoculture or sequential culture. Zygotes from hybrid F1 B6D2 mice were cultured in vitro to the blastocyst stage in Potassium (K(+)) simplex optimised medium (KSOM) throughout or in KSOM and switched to COOK blastocyst medium on day 3 (KSOM-CBM). Blastocysts were explanted on a feeder layer of mitomycin C-inactivated murine embryonic fibroblasts (MEF) in TX-WES medium for ES cell derivation. Sequential KSOM-CBM resulted in improved blastocyst formation compared to KSOM monoculture. ES cells were obtained from 32.1 of explanted blastocsyts cultured in KSOM-CBM versus 18.4 in KSOM alone. ES cell lines were characterized by morphology, expression of SSEA-1, Oct-4 and alkaline phosphatase activity, and normal karyotype. These results indicate that in vitro culture systems to produce blastocysts can influence the efficiency of ES cell line derivation.
Original languageEnglish
Pages (from-to)1017 - 1021
Number of pages5
JournalMolecular Reproduction and Development
Issue number8
Publication statusPublished - 2006
Externally publishedYes

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