Cryopreservation of chicken primordial germ cells by vitrification and slow freezing: A comparative study
| Autor: | Céline Tonus, Olivier Waroux, Benoît Vandenhove, Daniel Desmecht, Delphine Connan, Luc Grobet, Fabien Ectors, Jerome Wayet, Laurent Gillet, Nadine Antoine |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Time Factors Gonad Chick Embryo Biology Cryopreservation Flow cytometry Andrology 03 medical and health sciences Food Animals Freezing Botany medicine Animals Germ Vitrification Small Animals Slow freezing medicine.diagnostic_test Equine Embryo Flow Cytometry Embryonic stem cell Germ Cells 030104 developmental biology medicine.anatomical_structure Animal Science and Zoology |
| Zdroj: | Theriogenology. 88:197-206 |
| ISSN: | 0093-691X |
| DOI: | 10.1016/j.theriogenology.2016.09.022 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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