Dynamic medium containing growth differentiation factor-9 and FSH maintains survival and promotes in vitro growth of caprine preantral follicles after long-term in vitro culture.
The aim of the present study was to evaluate the effects of growth differentiation factor 9 (GDF-9) and FSH on the in vitro development of caprine preantral follicles cultured for 16 days. Ovarian fragments were cultured in αMEM⁺ (α-minimum essential medium, pH 7.2-7.4, 10 μg mL⁻¹ insulin, 5.5 μg mL⁻¹ transferrin, 5.0 ng mL⁻¹ selenium, 2 mM glutamine, 2 mM hypoxanthine and 1.25 mg mL⁻¹ bovine serum albumin) in the absence or presence of 200 ng mL⁻¹ GDF-9 and/or 50 ng mL⁻¹ FSH added during the first (Days 0-8) and/or second (Days 8-16) half of the culture period. Non-cultured and cultured fragments were processed for histological and ultrastructural analyses. After 16 days, all treatments using GDF-9 or FSH showed higher rates of follicular survival compared with αMEM⁺ alone. Compared with non-cultured control, sequential culture media containing GDF-9 and/or FSH significantly increased the percentage of developing follicles and follicle diameter. Moreover, a progressive increase in oocyte diameter was observed only with sequential culture medium containing GDF-9 until Day 8 followed by FSH (GDF-9/FSH) in the second half of the culture period. After 16 days of culture, ultrastructural analysis confirmed the integrity of follicles cultured in the presence of GDF-9/FSH. In conclusion, a dynamic medium containing GDF-9 and FSH (GDF-9/FSH) maintained follicular integrity and promoted activation of primordial follicles and growth during long-term in vitro culture of goat preantral follicles.
Laboratory of Manipulation of Oocytes and Preantral Follicles LAMOFOPA, Faculty of Veterinary Medicine, State University of Ceará, Av. Paranjana 1700, Campus Itaperi, 60740-903, Fortaleza, CE, Brazil. anelise_alves@yahoo.com.br
A M C V
AM
Alves
R N
RN
Chaves
R M P
RM
Rocha
L F
LF
Lima
P M
PM
Andrade
C A P
CA
Lopes
C E A
CE
Souza
A A A
AA
Moura
C C
CC
Campello
S N
SN
Báo
J
J
Smitz
J R
JR
Figueiredo
D000003:Abattoirs; D000818:Animals; D001938:Brazil; D002417:Cattle; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003433:Crosses, Genetic; D005260:Female; D005300:Fertility Agents, Female / Q000494:pharmacology*; D005640:Follicle Stimulating Hormone / Q000235:genetics / Q000494:pharmacology*; D006041:Goats / Q000502:physiology*; D055429:Growth Differentiation Factor 9 / Q000235:genetics / Q000494:pharmacology*; D006801:Humans; D059471:In Vitro Oocyte Maturation Techniques / Q000662:veterinary*; D009865:Oocytes / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure; D009866:Oogenesis / Q000187:drug effects; D006080:Ovarian Follicle / Q000187:drug effects* / Q000502:physiology / Q000648:ultrastructure; D011994:Recombinant Proteins / Q000494:pharmacology; D013997:Time Factors; D046509:Tissue Culture Techniques / Q000662:veterinary
D005300:Fertility Agents, Female; C525897:GDF9 protein, human; D055429:Growth Differentiation Factor 9; D011994:Recombinant Proteins; D005640:Follicle Stimulating Hormone