Effect of sequential medium on in vitro culture of goat ovarian cortical tissue.
A sequential medium was evaluated on the survival, activation and growth rates of caprine preantral follicles submitted to a long-term culture period, aiming to establish an ideal in vitro culture system. Ovarian fragments were cultured for 16 days in α-MEM(+) alone or supplemented with hormones (GH and/or FSH) added sequentially on different days of culture. Ovarian fragments were cultured in the first (days 0-8) and second (days 8-16) halves of the culture period, generating 10 treatments: α-MEM(+)/α-MEM(+), FSH/FSH, FSH/GH, FSH/FSH+GH, GH/GH, GH/FSH, GH/FSH+GH, FSH+GH/FSH+GH, FSH+GH/FSH and FSH+GH/GH. Follicle morphology, viability and ultrastructure were analyzed. After day 1 of culture, FSH treatments maintained the percentage of normal follicles similar to the fresh control. At day 16 of culture, the treatment FSH/GH showed the highest (P<0.05) percentage of normal follicles. The ultrastructure of follicles was preserved in the fresh control and FSH/GH treatment. Follicles cultured with FSH/GH had a higher (P<0.05) viability than α-MEM(+); however the viability was lower (P<0.05) when compared to the fresh control. The FSH/GH treatment showed the highest (P<0.05) percentage of follicular activation and secondary follicle formation and produced the largest (P<0.05) mean follicular diameter after 16 days of culture. In conclusion, a sequential medium supplemented with FSH followed by GH during a long-term culture maintains the survival, viability and ultrastructure of goat preantral follicles, and promotes activation and secondary follicles.
Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), Veterinary Faculty, State University of Ceará, Av. Paranjana, 1700, Campus do Itaperi, Fortaleza, 60740-903, CE, Brazil. dmmvet@hotmail.com
D M
DM
Magalhães-Padilha
P M
PM
Andrade
E T
ET
Sales
V R
VR
Araujo
I M T
IM
Lima
S V
SV
Castro
L R
LR
Faustino
C A P
CA
Lopes
C C
CC
Campello
S N
SN
Báo
M O
MO
Gastal
E L
EL
Gastal
J R
JR
Figueiredo
D000818:Animals; D003470:Culture Media; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology; D006041:Goats / Q000502:physiology*; D013006:Growth Hormone / Q000494:pharmacology; D010053:Ovary / Q000187:drug effects* / Q000502:physiology* / Q000648:ultrastructure; D013997:Time Factors; D046509:Tissue Culture Techniques / Q000662:veterinary*
D003470:Culture Media; D005640:Follicle Stimulating Hormone; D013006:Growth Hormone