Le module SPF a pour mission d’héberger et reproduire les lignées transgéniques et de garantir le statut sanitaire « Specific Pathogen Free ».
Ce niveau sanitaire est défini par rapport à une liste de référence d’organismes pathogènes viraux, bactériens ou parasitaires à exclure des élevages. Ainsi, l’absence de ces pathogènes assure le bien-être des animaux, la qualité de la reproduction et la fiabilité des résultats expérimentaux. Le maintien de ce statut préserve les élevages de catastrophes sanitaires pouvant avoir des impacts scientifiques et financiers importants. Un autre avantage du statut sanitaire SPF est la facilitation des échanges de lignées transgéniques pour des collaborations scientifiques nationales et internationales. En effet, la délivrance de certificats statuts sanitaires SPF accélère les démarches administratives et les acceptations d’échange auprès des services vétérinaires, douaniers et responsables d’animaleries. La plateforme animalière SPF a également pour mission de cryoconserver les lignées de souris, transgéniques principalement.
Prestations
Hébergement et gestion de lignée
Le module SPF vous propose de conserver vos lignées sous forme respirante au statut sanitaire SPF. Le propriétaire de la lignée gère ces colonies par l’intermédiaire du logiciel de gestion Mayakind. Le travail de suivi du bien-être animal et de la gestion des lignées (identification, biopsie, sevrage) est assuré par l’équipe de 5 zootechniciens. Annuellement 800 croisements sont lancés, 12 000 souriceaux sont ainsi générés. Près de 2500 souris sont sorties annuellement du module SPF pour expérimentation dans les différents centres de recherche de Lyon.
Transfert aseptique d’embryon
Pour que les lignées n’ayant pas le statut SPF puissent accéder à la plateforme SPF, il faut « décontaminer » ces lignées par une technique d’embryologie appelée « transfert aseptique d’embryons». Nous demandons de nous fournir 3-5 mâles qui seront hébergés au module 2 d’ALECS qui n’a aucun contact avec le module SPF. Seules les femelles superovulées seront mises en contact pendant une nuit. La suite de la technique consiste à nettoyer les embryons fécondés par bains successifs, puis de les réimplanter par microchirurgie au niveau de l’ovaire de mères porteuses dans des conditions aseptiques au niveau des expérimentales du SPF. Un contrôle sanitaire est réalisé afin de confirmer l’élimination des pathogènes sur les souriceaux obtenus.
Cryopréservation des lignées
La cryopréservation est l’ensemble des techniques qui consistent à la congélation des gamètes des lignées de souris transgéniques dans l’azote et à la reviviscence des souris. Ces savoir-faire permettent la « sauvegarde » de lignées. Elles sont ainsi protégées de la dérive génétique et peuvent être revivifiées à tout moment en cas de problèmes sanitaires ou de problème de stérilité. De plus, avec le renforcement de la réglementation du transport des animaux et des difficultés qui en découlent, l’échange de souches via du matériel congelé est une solution de plus en plus employée. Enfin, la cryoconservation permet de réduire le nombre de lignées vivantes, ce qui est en parfait accord avec la règle de réduction des animaux.
Cryopréservation de sperme
Nous proposons la cryopréservation de sperme via le Jax Sperm Cryo Kit. Pour cela, il faut nous fournir deux mâles de 10-16 semaines le jour des prélèvements. Nous nous occupons de récupérer les spermatozoïdes dans les milieux appropriés du kit, de réaliser 20 paillettes et de les congeler selon le protocole. Les paillettes sont ensuite envoyées via un dry-shipper sur le site de Janvier Labs. Un contrôle qualité (Fécondation In Vitro) est alors réalisé par la société Janvier Labs sur une des paillettes envoyées sur le fonds génétique que vous préciserez. Le pourcentage de passage à deux cellules valide la qualité des paillettes. Le stock se fait par la société Janvier Labs (environ 100 €/an/lignée à votre charge) sur deux sites sécurisés. Vous pouvez également choisir de stocker vos paillettes dans vos propres bacs à azote.
Fécondation In Vitro
La reviviscence de lignée à partir de paillettes congelées est une prestation récemment développée dans notre service. Merci de nous consulter pour les conditions de réalisation.
Présentation du bâtiment et statut sanitaire
La plateforme a été conçue et fonctionne pour maintenir le statut SPF :
filtration de l’air entrant par la centrale de traitement d’air ;
zone protégée en surpression instaurant un gradient de pression décroissant des zones les plus propres à la laverie ;
stérilisation de tous les produits entrants (eau, aliment, litière, cage…) par autoclave double entrée, sas à désinfection gazeuse ou filtration ;
procédures strictes concernant le circuit des déchets et le ménage ;
procédures particulières concernant le personnel entrant en zone protégée (douche à chaque entrée, vêtements stériles changés quotidiennement, port d’un masque et d’une charlotte, changement fréquent des gants) ;
achat d’animaux au statut sanitaire SPF, livrés par les fournisseurs agréés (Charles River, Harlan…).
Un contrôle sanitaire (bactériologie, virologie, parasitologie) des animaux sentinelles est réalisé tous les trimestres afin de vérifier le maintien du statut SPF de la plateforme.
Contact
Bibliographie
Voici les derniers articles déclarés pour la plateforme ALECS SPF :
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