Production of functional gametes from cryopreserved primordial germ cells of the Japanese quail.
The Japanese quail (Coturnix japonica) is a valuable bird as both an experimental animal, for a wide range of scientific disciplines, and an agricultural animal, for the production of eggs and meat. Cryopreservation of PGCs would be a feasible strategy for the conservation of both male and female fertility cells in Japanese quail. However, the effects of freeze-thaw treatment on viability, migration ability and germline transmission ability of quail PGCs still remain unclear. In the present study, male and female PGCs were isolated from the blood of 2-day-old embryos, which were cooled by slow freezing and then cryopreserved at -196 C for 77-185 days, respectively. The average recovery rate of PGCs after freeze-thawing was 47.0%. The viability of PGCs in the frozen group was significantly lower than that of the control group (P<0.05) (85.5% vs. 95.1%). Both fresh and Frozen-thawed PGCs that were intravascularly transplanted into recipient embryos migrated toward and were incorporated into recipient gonads, although the number of PGCs settled in the gonads was 48.5% lower in the frozen group than in the unfrozen control group (P<0.05). Genetic cross analysis revealed that one female and two male recipients produced live progeny derived from the frozen-thawed PGCs. The frequency of donor-derived offspring was slightly lower than that of unfrozen controls, but the difference was not significant (4.0 vs. 14.0%). These results revealed that freeze-thaw treatment causes a decrease in viability, migration ability and germline transmission ability of PGCs in quail.
Division of Germ Cell Biology, National Institute for Basic Biology, National Institute of Natural Sciences, Aichi 444-8787, Japan.
Yoshiaki
Y
Nakamura
Mariko
M
Tasai
Kumiko
K
Takeda
Keijiro
K
Nirasawa
Takahiro
T
Tagami
D000818:Animals; D018070:Biological Specimen Banks; D002465:Cell Movement; D002470:Cell Survival; D003370:Coturnix / Q000196:embryology / Q000235:genetics / Q000502:physiology*; D015925:Cryopreservation / Q000662:veterinary*; D015536:Down-Regulation; D046149:Embryo Culture Techniques / Q000662:veterinary; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology* / Q000637:transplantation; D056727:Endangered Species; D005240:Feasibility Studies; D005260:Female; D005298:Fertility; D007564:Japan; D008297:Male; D009154:Mutation; D010063:Ovum / Q000166:cytology / Q000502:physiology* / Q000637:transplantation; D013094:Spermatozoa / Q000166:cytology / Q000502:physiology* / Q000637:transplantation; D033581:Stem Cell Transplantation; D018183:Transplantation Chimera / Q000235:genetics / Q000502:physiology*