Service de Cytogénétique & d’Embryologie Hôpital Necker-Enfants Malades M Vekemans The Necker Human Embryo Resource Foetal pathology Embryology Molecular Genetics G Goudefroye H Etchevers S Thomas C Babarit T Attié-Bitach F Encha-Razavi M Bonnière C Esculpavit J Martinovic

Embryo Foetus New- born “ Pre”- embryo Fertilization D1 D15 WG10 WG40 D8 Birth Inner cell mass Preimplantation genetic diagnosis Pluripotent stem cells Embryonic and foetal cells / tissues Gene/protein expression Multipotent (stem) cells IVF Elective abortion Spontaneous abortion Medical terminations

Preimplantation genetic diagnosis

Pluripotent stem cells

Gene/protein expression

Multipotent (stem) cells

Mostly RU-486 Occasionally aspiration Embryos < 50 days 5-9 wg C9-C19 Late embryos/early foetuses 8-12 wg C18-23+ whole embryonic sac 60% recovery entire / pieces fragments organs, parts never whole brain Techniques for elective abortion Whole embryo with trophoblast Pieces (spinal cord)

http://embryology.med.unsw.edu.au/embryo.html 2 wg 3-4 wg 9-10.5 wg fetus 5-9 wg

http://embryology.med.unsw.edu.au/embryo.html

Distribution of embryo ages

Information database Anonymous identifier at our end only independent abortion clinic can link to patient has been useful once for HIV testing a requirement of bioethics law Transport conditions: PBS, RPMI, delays State, our staging of embryo Karyotype and/or anomalies Final use: fixation (IHC, cold/hot ISH) cell culture to derive migratory neural crest cells extraction for RNA fixation for IHC microdissection expression studies enzyme complex activities storage and use of individual slides and tubes

Anonymous identifier at our end

only independent abortion clinic can link to patient

has been useful once for HIV testing

a requirement of bioethics law

Transport conditions: PBS, RPMI, delays

State, our staging of embryo

Karyotype and/or anomalies

Final use:

fixation (IHC, cold/hot ISH)

cell culture to derive migratory neural crest cells

extraction for RNA

fixation for IHC

microdissection

expression studies

enzyme complex activities

storage and use of individual slides and tubes

Gene expression: in situ hybridization FGF10 , C19 NANOG , C15

Gene expression: SAGE and microarrays

Gene expression: (q)RT-PCR

Gene translation: immunohistochemistry Synaptogamin (brown) in intestine Neural cell adhesion molecule (white) in nerves and spinal cord

Near-future research avenues Tissue-specific ch romatin i mmuno p recipitation ChIP-sequencing Many other possibilities miRNAs, splice variants…

Tissue-specific ch romatin i mmuno p recipitation

ChIP-sequencing

Many other possibilities

miRNAs, splice variants…

Near-future research avenues Cell line derivation Functional studies in vitro Transplantation and function in vivo N-CAM (green), hNCC Fetal pericytes (green) in NOD-SCID mouse model (Crisan et al. 2008)

Cell line derivation

Functional studies in vitro

Transplantation and function in vivo

© Nick Kim, http://nearingzero.net/