Preparation of rat serum suitable for mammalian whole embryo culture

Masanori Takahashi, Sayaka Makino, Takako Kikkawa, Noriko Osumi

Research output: Contribution to journalArticlepeer-review

15 Citations (Scopus)

Abstract

Mammalian whole embryo culture (WEC) is a widely used technique for examining pharmacological toxicity in developing mouse and rat embryos and for investigating the mechanisms of developmental processes. Immediately centrifuged (IC) rat serum is commonly used for WEC and is essential for the growth and development of cultured mouse and rat embryos ex vivo. For the culture of midgestation embryos (i.e., E8.0-12.5 for the mouse, and E10.0-14.5 for the rat), 100% rat serum is the best media for supporting the growth of the embryo ex vivo. To prepare rat serum suitable for WEC, the collected blood should be centrifuged immediately to separate the blood cells from the plasma fraction. After centrifugation, the fibrin clot forms in the upper layer; this clot should be squeezed gently using a pair of sterile forceps and subsequently centrifuged to completely separate the blood cells from the serum. In this video article, we demonstrate our standard protocol for the preparation of optimal IC rat serum, including blood collection from the abdominal aorta of male rats and extraction of the serum by centrifugation.

Original languageEnglish
Article numbere51969
JournalJournal of Visualized Experiments
Issue number90
DOIs
Publication statusPublished - 2014 Mar 8

Keywords

  • Abdominal aorta
  • Fibrin clot
  • Hemolysis
  • Issue 90
  • Male rats
  • Physiology
  • Rat serum
  • WEC

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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