In vitro growth and maturation of isolated caprine preantral follicles: Influence of insulin and FSH concentration, culture dish, coculture, and oocyte size on meiotic resumption.
The aims of this study were: (1) to evaluate the effect of different insulin concentrations, alone or in combination with either a fixed FSH concentration or increasing FSH concentrations on the in vitro culture of isolated caprine preantral follicles and (2) to analyze the efficiency of two IVM media and maturation culture systems (with or without coculture with in vivo grown oocytes) on the meiosis resumption. Secondary follicles were cultured for 18 days in a basic medium supplemented with low- or high-insulin concentration alone or with a fixed FSH concentration or with increasing FSH concentrations. Oocytes grown in vivo or in vitro were matured alone or cocultured. The high-insulin concentration associated with fixed FSH treatment had higher meiotic resumption rate (P < 0.05) and was the only treatment capable of producing oocytes in metaphase II. The rates of germinal vesicle, germinal vesicle breakdown, metaphase I, metaphase II (MII), meiotic resumption, and oocyte diameter were similar between the maturation media. In conclusion, a basic medium supplemented with 10-μg/mL insulin and 100-μg/mL FSH throughout the culture period improved meiotic resumption rate and produced MII oocytes from caprine preantral follicles cultured in vitro. The MII rate was similar between in vivo and in vitro grown oocytes ≥110 μm.
Laboratory of Manipulation of Oocyte and Preantral Follicles (LAMOFOPA), Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil.
G M
GM
Silva
Laboratory of Manipulation of Oocyte and Preantral Follicles (LAMOFOPA), Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil.
I R
IR
Brito
Laboratory of Manipulation of Oocyte and Preantral Follicles (LAMOFOPA), Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil.
A D
AD
Sales
Laboratory of Manipulation of Oocyte and Preantral Follicles (LAMOFOPA), Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil.
F L N
FLN
Aguiar
Laboratory of Manipulation of Oocyte and Preantral Follicles (LAMOFOPA), Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil.
A B G
ABG
Duarte
Laboratory of Manipulation of Oocyte and Preantral Follicles (LAMOFOPA), Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil.
V R
VR
Araújo
Laboratory of Manipulation of Oocyte and Preantral Follicles (LAMOFOPA), Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil.
L A
LA
Vieira
Postgraduate Biotechnology, Potiguar University/Laureate International Universities, Natal, CE, Brazil.
D M
DM
Magalhães-Padilha
Laboratory of Manipulation of Oocyte and Preantral Follicles (LAMOFOPA), Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil.
L F
LF
Lima
Laboratory of Manipulation of Oocyte and Preantral Follicles (LAMOFOPA), Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil.
B G
BG
Alves
Human Reproduction Section, Division of Urology, Department of Surgery, Sao Paulo Federal University, Sao Paulo, Brazil.
L B R
LBR
Silveira
Human Reproduction Section, Division of Urology, Department of Surgery, Sao Paulo Federal University, Sao Paulo, Brazil.
E G
EG
Lo Turco
Laboratory of Manipulation of Oocyte and Preantral Follicles (LAMOFOPA), Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil.
A P
AP
Rodrigues
Laboratory of Manipulation of Oocyte and Preantral Follicles (LAMOFOPA), Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil.
C C
CC
Campello
Department of Animal Sciences, University of Illinois, Urbana-Champaign, Urbana, IL, USA.
M B
MB
Wheeler
Laboratory of Manipulation of Oocyte and Preantral Follicles (LAMOFOPA), Faculty of Veterinary Medicine, State University of Ceará, Fortaleza, CE, Brazil. Electronic address: jrf.lamofopapapers@gmail.com.
J R
JR
Figueiredo
D000818:Animals; D018920:Coculture Techniques / Q000662:veterinary*; D003470:Culture Media; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D006041:Goats; D059471:In Vitro Oocyte Maturation Techniques / Q000379:methods / Q000662:veterinary*; D007328:Insulin / Q000494:pharmacology*; D008540:Meiosis; D009865:Oocytes / Q000254:growth & development*; D006080:Ovarian Follicle / Q000187:drug effects*
D003470:Culture Media; D007328:Insulin; D005640:Follicle Stimulating Hormone
Embryo corral; Oocyte competence; Secondary follicles