title: Follicle development in cryopreserved bitch ovarian tissue grafted to immunodeficient mouse.
abstract: The present study evaluated: (1) in vivo follicular development in canine ovarian tissue after slow freezing and xenotransplantation; and (2) the use of erythropoietin (EPO) as an angiogenic factor to optimise the transplantation procedure. Frozen-thawed ovarian tissue from five bitches was grafted into severe combined immunodeficient (SCID) mice (n=47) treated with or without EPO (500 IU kg(-1), once daily for 3 days) (Groups A and B, respectively) and analysed after 0, 1, 8 or 16 weeks. Follicle grade, follicle density, follicle morphology and stromal cells density were assessed by histological analysis, whereas vascularisation of the graft was quantified by immunohistochemistry with anti-α-smooth muscle actin antibody. Despite a massive loss of follicles after grafting, secondary follicle density was higher at 8 and 16 weeks than at 1 week regardless of EPO treatment. EPO significantly improved early follicle morphology and stromal cell density after 8 weeks and blood vessel density at 16 weeks after transplantation (P<0.05). Intact secondary follicles with more than three granulosa cells layers were observed 16 weeks after transplantation. The results suggest that canine ovarian tissue can be successfully preserved by our slow-freezing protocol because the tissue showed follicular growth after xenotransplantation. EPO treatment did not lessen the massive loss of follicles after transplantation.
authors.affiliation: Equipe Cryobio, Unité Propre de Soutien de Programme Interaction Cellule Environnement, VetAgroSup/ISARA Lyon, Université de Lyon, VetAgro Sup, 2011.03.101, UPSP I.C.E.-Equipe CRYOBIO, F-69280 Marcy l'Etoile, France. l.commin@vetagro-sup.fr
authors.forename: L
authors.initials: L
authors.lastname: Commin
authors.forename: S
authors.initials: S
authors.lastname: Buff
authors.forename: E
authors.initials: E
authors.lastname: Rosset
authors.forename: C
authors.initials: C
authors.lastname: Galet
authors.forename: A
authors.initials: A
authors.lastname: Allard
authors.forename: P
authors.initials: P
authors.lastname: Bruyere
authors.forename: T
authors.initials: T
authors.lastname: Joly
authors.forename: P
authors.initials: P
authors.lastname: Guérin
authors.forename: V
authors.initials: V
authors.lastname: Neto
mesh_terms: D000818:Animals; D002452:Cell Count; D015925:Cryopreservation / Q000662:veterinary; D004285:Dogs; D005260:Female; D012739:Gonadal Steroid Hormones / Q000097:blood; D016867:Immunocompromised Host / Q000502:physiology; D051379:Mice; D016513:Mice, SCID; D006080:Ovarian Follicle / Q000166:cytology / Q000502:physiology* / Q000637:transplantation*; D010053:Ovary / Q000166:cytology / Q000637:transplantation; D013997:Time Factors; D014183:Transplantation, Heterologous / Q000276:immunology; D014621:Vagina / Q000166:cytology / Q000378:metabolism
chemical_list: D012739:Gonadal Steroid Hormones