Mammalian unit

Methods

Bovine in vitro embryo production (from slaughterhouse material)

Porcine in vitro embryo production (from slaughterhouse material)

Feline in vitro embryo production (from discarded ovaries/testes after standard spaying in local veterinary clinics)

In vitro embryo production of ovines and other species can in some instances be set up

For all species development is followed:

• Cumulus expansion after maturation

• Cleavage after fertilization

• Embryo development to blastocyst stage (blastocyst quality grading and blastocyst developmental stage)

Further work with blastocysts or earlier developmental stages:

• Individual snap freezing in LN2 and storage in -80℃ for e.g. RNA-analysis (see below).

• Individual fluorescent staining for confocal or fluorescence microscopy. Protocols currently available:

  • Neutral lipids (LipidTox or Nile Red)

  • Active mitochondria (Mitotrackers)

  • F-Actin (Alexafluor Phalloidin)

  • Nuclei (DAPI, Hoechst or Draq5)

Connected services:

1. Ovum Pic-Up (bovines)

2. RNA-expression analyses of embryos from -80℃ storage

3. Epigenetic analyses of embryos from -80℃ storage

4. Embryo transfer

5. Analyses of culture media

6. Hazard analysis