Journal
ANALYTICAL BIOCHEMISTRY
Volume 416, Issue 2, Pages 228-230Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2011.05.033
Keywords
Differential staining; Apoptosis; Preimplantation embryo; Immunofluorescence
Funding
- Fonds voor wetenschappelijk onderzoek Vlaanderen (FWO Vlaanderen)
- Agentschap voor innovatie door wetenschap en technologie (IWT)
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Several approaches have been described for differential staining of blastocysts, but these methods are often time-consuming and unreliable. Here we describe a method for simultaneous differential staining and detection of apoptosis. The differential staining is based on the transcription factor CDX2 which is localized in the nucleus of trophectoderm (TE) cells but absent in the inner cell mass (ICM). Apoptosis is detected by staining of active caspase-3, a key player in several apoptotic pathways. This new approach represents a robust method for quantifying simultaneously ICM/TE ratio and apoptotic cell ratio in bovine, murine, porcine, and human blastocysts. (C) 2011 Elsevier Inc. All rights reserved.
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