title: Cryopreservation of chicken primordial germ cells by vitrification and slow freezing: A comparative study.
abstract: In the present study, we compare a classical slow freezing (SLF) method and an aseptic vitrification (Vitrif) technique to cryopreserve a stable primordial germ cell (PGCs) line issued from the Ardennaise chicken breed. Viability immediately after warming was close to 80% and did not differ between the two cryopreservation methods. Proliferation tended to be slower for both cryopreservation methods compared with controls, but the difference was significant only for Vitrif. No difference was found between the two methods after flow cytometry analysis of stage-specific embryonic antigen-1 expression and reverse transcription-polymerase chain reaction on several factors related to PGC phenotype. After 1 week in culture, all cryopreserved cells reached controls' main morphologic and expanding (viability/proliferation) features. However, SLF generated more unwanted cells clusters than Vitrif. After injection of the PGCs into recipient embryos, vitrified PGCs reported a clear, yet not significant, tendency to colonize the gonad at a higher rate than slow frozen PGCs. SLF in cryovials remains simple, inexpensive, and less technically demanding than Vitrif. Nevertheless, the intrinsic advantages of our aseptic Vitrif method and the present study suggest that this should be considered as safer than classical SLF for cryopreserving chicken PGCs.
authors.affiliation: Embryology Unit, FARAH (Fundamental and Applied Research for Animals and Health) Research Center and Faculty of Veterinary Medicine, University of Liège, Liège, Belgium.
authors.forename: C
authors.initials: C
authors.lastname: Tonus
authors.affiliation: Embryology Unit, FARAH (Fundamental and Applied Research for Animals and Health) Research Center and Faculty of Veterinary Medicine, University of Liège, Liège, Belgium.
authors.forename: D
authors.initials: D
authors.lastname: Connan
authors.affiliation: Embryology Unit, FARAH (Fundamental and Applied Research for Animals and Health) Research Center and Faculty of Veterinary Medicine, University of Liège, Liège, Belgium; Histology Unit, FARAH Research Center and Faculty of Veterinary Medicine, University of Liège, Liège, Belgium.
authors.forename: O
authors.initials: O
authors.lastname: Waroux
authors.affiliation: Embryology Unit, FARAH (Fundamental and Applied Research for Animals and Health) Research Center and Faculty of Veterinary Medicine, University of Liège, Liège, Belgium.
authors.forename: B
authors.initials: B
authors.lastname: Vandenhove
authors.affiliation: Embryology Unit, FARAH (Fundamental and Applied Research for Animals and Health) Research Center and Faculty of Veterinary Medicine, University of Liège, Liège, Belgium.
authors.forename: J
authors.initials: J
authors.lastname: Wayet
authors.affiliation: Immunology and Vaccinology Unit, FARAH Research Center and Faculty of Veterinary Medicine, University of Liège, Liège, Belgium.
authors.forename: L
authors.initials: L
authors.lastname: Gillet
authors.affiliation: Pathology Unit, FARAH Research Center and Faculty of Veterinary Medicine, University of Liège, Liège, Belgium.
authors.forename: D
authors.initials: D
authors.lastname: Desmecht
authors.affiliation: Histology Unit, FARAH Research Center and Faculty of Veterinary Medicine, University of Liège, Liège, Belgium.
authors.forename: N
authors.initials: N
authors.lastname: Antoine
authors.affiliation: Mammalian Transgenics Platforms, FARAH and GIGA (Grappe Interdisciplinaire de Génoprotéomique Appliquée) Research Centers, University of Liège, Liège, Belgium.
authors.forename: F J
authors.initials: FJ
authors.lastname: Ectors
authors.affiliation: Embryology Unit, FARAH (Fundamental and Applied Research for Animals and Health) Research Center and Faculty of Veterinary Medicine, University of Liège, Liège, Belgium. Electronic address: lgrobet@ulg.ac.be.
authors.forename: L
authors.initials: L
authors.lastname: Grobet
mesh_terms: D000818:Animals; D002642:Chick Embryo; D015925:Cryopreservation / Q000662:veterinary*; D005434:Flow Cytometry / Q000662:veterinary; D005615:Freezing; D005854:Germ Cells / Q000187:drug effects / Q000502:physiology*; D013997:Time Factors; D058989:Vitrification
keywords: Aseptic vitrification; Chicken primordial germ cells; Cryopreservation; Genetic resources preservation; Male and female germplasm; Slow freezing