Isolation and culture of chicken primordial follicles.
The establishment of a primordial follicle culture system is important for the study of follicular development. Hence, the objective of this study was to isolate chicken primordial follicles and establish culture methods. Ovaries from 2-wk-old chickens were treated with trypsin-EDTA, collagenase II, or collagenase type IA, along with a mechanical isolation technique. Isolated follicles were cultured under different conditions. Results showed a significant difference in the follicular recovery and survival rates among different enzymes and methods used. The maximal follicular yield was obtained by trypsin+EDTA and collagenase II digestion, followed by collagenase type IA digestion. However, the highest follicular viability rate was observed in groups of collagenase type IA digestion and the mechanical isolation method. Enzymatic treatment resulted in higher misshapen oocytes or follicles, though the diameters of the follicles were not significantly changed. In addition, our follicle culture results for different conditions showed maximal survival rates of primordial follicles in alginate hydrogel beads after 12 d of culture. Thus, we successfully established methods for isolating and culturing chicken primordial follicles. The present method will greatly facilitate investigation of the regulation of follicular development.
Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310058, P. R. China Department of Poultry Husbandry, Sindh Agriculture University, Tandojam 760060, Pakistan.
Imdad Hussain
IH
Leghari
Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310058, P. R. China.
Dan
D
Zhao
Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310058, P. R. China.
Yuling
Y
Mi
Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310058, P. R. China cqzhang@zju.edu.cn.
Caiqiao
C
Zhang
D000818:Animals; D018929:Cell Culture Techniques / Q000662:veterinary; D002645:Chickens / Q000254:growth & development*; D005260:Female; D006080:Ovarian Follicle / Q000166:cytology* / Q000254:growth & development*
chicken; culture; isolation; primordial follicle