Canine preantral follicles cultured with various concentrations of follicle-stimulating hormone (FSH).
The objective was to evaluate the effects of various concentrations of exogenous FSH during in vitro culture of isolated canine preantral follicles. Preantral secondary follicles (>200 microm) were isolated by microdissection and cultured for 18 d in supplemented alpha-Minimum Essential Medium (alpha-MEM). There were three treatment groups: 1) absence of FSH (control medium); 2) FSH100 (fixed concentration of 100 ng/mL throughout the entire culture period); and 3) sequential FSH (FSHSeq - 100, 500, and 1,000 ng/mL were added sequentially). Following culture, all follicles from all treatments were still viable (marked green by calcein-AM). The initial (D0) average follicle diameter for the control, FSH100, and FSHSeq was (mean +/- SEM) 298.96 +/- 7.02, 286.00 +/- 5.87, and 275.39 +/- 174 6.55 microm, respectively (P > 0.05). Mean diameter of follicles treated with FSHSeq on Day 18 (D18-439.80 +/- 14.08 microm) was greater than those of the other treatments (P < 0.05). Daily follicular growth rate (microm/d) of follicles in the FSHSeq treatment (6.47 +/- 0.55) was significantly faster than for both the control (3.67 +/- 0.32) and FSH100 (4.47 +/- 0.38) treatments. Furthermore, FSH100 and FSHSeq treatments had a significantly higher rate of antrum formation than the control group on D12 of culture, whereas after D12, FSH100 had a significantly higher rate of extrusion compared to the control (P < 0.05). In conclusion, the sequential addition of FSH to the culture medium maintained the survival of isolated canine preantral follicles and promoted an increased rate of follicular growth and antrum formation.
Faculty of Veterinary Medicine, Laboratory of Carnivore Reproduction (LRC), State University of Ceara, Fortaleza, CE, Brazil. michellekarenbs@hotmail.com
Michelle Karen Brasil
MK
Serafim
Valdevane Rocha
VR
Araújo
Gerlane Modesto
GM
Silva
Ana Beatriz Graça
AB
Duarte
Anderson Pinto
AP
Almeida
Roberta Nogueira
RN
Chaves
Cláudio Cabral
CC
Campello
Cláudio Afonso Pinho
CA
Lopes
José Ricardo
JR
de Figueiredo
Lúcia Daniel Machado
LD
da Silva
D000818:Animals; D003470:Culture Media; D004285:Dogs; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D006080:Ovarian Follicle / Q000033:anatomy & histology / Q000187:drug effects* / Q000254:growth & development; D046509:Tissue Culture Techniques / Q000662:veterinary*
D003470:Culture Media; D005640:Follicle Stimulating Hormone